1.Cloning and expression of urease B subunit (UreB) of Helicobacter pylori
Chao WU ; Ning WANG ; Xiaopeng YUAN
Chinese Journal of Laboratory Medicine 2001;0(05):-
Objective To obtain the recombinant urease B subunit (ureB) of Helicobacter pylori (Hp) and apply it in serological detection of Hp infected patients. Methods Urease B subunit gene was amplified from the complete genome of Helicobacter pylori by PCR, and cloned into the PinPoint TM Xa Ⅲ fusion expression vector, then was sequenced. The protein of urease B subunit was expressed in E.coli JM109 and purified with affinity chromatography. 113 serum samples of peptic ulcer patients were detected by ELISA combined with purified UreB protein. Results DNA sequence analysis showed the nucleic acid sequence homology of ureB gene was 96.44% and the putative amino acid sequence homology was 99.65%. The recombinant UreB protein was composed of 571 amino acid residues and kept original immunologic reaction with corresponding antibody, and its purity was over 90% after affinity chromatography. The results of ELISA associated with recombination UreB antigen showed the sensitivity and specificity was 92%, 98.5%. Conclusion The recombinant UreB protein will be of value for clinical serodiagnosis and epidemiological study of Helicobacter pylori.
2.Construction and identification of adenovirus-recombined vector of CTLA4Ig-IRES2-I?B?
Ning LI ; Yukang YUAN ; Ju WU
Journal of Third Military Medical University 1984;0(02):-
Objective To construct adenovirus vector harboring CTLA4Ig-IRES2-I?B? gene for the study of enhancement blockade of T cell costimulatory pathway. Methods The IRES2 fragment was cloned in to the shuttle plasmid pAdtrack-CMV-CTLA4Ig, and then the human I?B? fragment was constructed with the link of the IRES2 to the pAdtrack-CMV-CTLA4Ig. Subsequently, adenovirus vector harboring CTLA4Ig-IRES2-I?B? was constructed by homologous recombination in E. coli BJ5183, and recombinant vector was packaged and propagated in 293 cells. Results The recombinant CTLA4Ig-IRES2-I?B? adenovirus was constructed by homologous recombination and identified by PCR and restrictive enzyme digestion methods. Conclusion The recombinant CTLA4Ig-IRES2-I?B? adenovirus may be used as a novel immunosuppressive agent for gene therapy in organ transplantation.
3.Effect of hepatitis B immunoglobulin to prevent de novo hepatitis B infection after renal transplantation
Ning LI ; Xiaotong WU ; Mingjun WANG ; Wenping GUO ; Yuan DONG ; Zuan FAN ; Yuan NING ; Tingting LIU
Chinese Journal of Organ Transplantation 2012;33(2):105-108
Objective To summarize the safety and efficacy of low dose of hepatitis B immunoglobulin (HBIG) for prevention of de novo hepatitis B infection after renal transplantation.MethodsThe clinical data of 138 patients who received renal transplantation without hepatitis B infection between January 2007 and June 2010 were retrospectively studied (study group).All the patients in study group were given low dose of HBIG injection before transplantation.The HBsAb titer was monitored regularly after transplantation,and the dosage of HBIG adjusted according to the level of the HBsAb titer.HBIG was implied to all patients in the study group for more than one year.The clinical data of 196 patients who received renal transplantation without hepatitis B infection between January 2004 and December 2006 served as the control group.These 196 patients were not treated with HBIG.The incidence of de novo hepatitis B infection,and acute rejection of these two groups was analyzed.The one-year graft and patients survival rate was also investigated.Results During the follow- up period of 12 months,only one case in the study group had de novo hepatitis B infection (0.7%) 6 months after renal transplantation,while 11 cases (5.6%) in the control group had de novo hepatitis B infection,in which 2 cases were died from acute hepatic failure.The incidence of de novo hepatitis B infection had statistically difference between the two groups (P<0.05).The incidence of acute rejection in the study and control groups was 13.8% and 17.3% respectively (P>0.05).The one-year graft and patient survival rate in the study and control groups was 96.4% and 97.8%,and 90.3% and 91.8% respectively (P<0.05).ConclusionLow dose of HBIG is effective and safe for prevention of de novo hepatitis B infection after renal transplantation.
4.The effects of dexmedetomidine for different loading dose on blood pressure of SICU patients conscious-sedation derivative phase
Zhengying JIANG ; Guixin WU ; Ning ZHANG ; Jianguo YUAN
Chongqing Medicine 2014;(5):516-517,520
Objective To observe the effects of dexmedetomidine for different loading dose on blood pressure of surgery inten-sive care unit(SICU) patients conscious-sedation derivative phase .Methods 60 patients after general anaesthesia were divided into routine dose group(dexmedetomidine 1 .00 μg/kg ,group C) ,middle dose group(dexmedetomidine 0 .75 μg/kg ,group M ) and low dose group(dexmedetomidine 0 .50 μg/kg ,group L) ,and the loading time were reset 10 min .sedative effect was evaluated by Ram-say sedation score .Record the mean arterial blood pressure of everyone before administration ,5 min after ,15 min after ,30 min af-ter ,60 min after .Results All patients achieved good conscious-sedation .Compared with pro-administration ,the mean arterial blood pressure of group C descend significantly .The mean arterial blood pressure of group M and L descend significantly ,and then they were recover over time .60 min after the treatment ,the mean arterial blood pressure in group C was significantly lower than group L and goup M (P<0 .05) .Conclusion Small dose dexmedetomidine can achieved good conscious-sedation for SICU patients .It has less influence on hemodynamics .
5.Effect of pneumatic lithotripsy treatment under ureteroscopic on urinary catheter encrustation
Chunlin LIU ; Ning LIU ; Zongshan WU ; Yuan QU
Clinical Medicine of China 2014;30(2):208-210
Objective To evaluate the therapy effect of pneumatic lithotripsy treatment under ureteroscopic on urinary catheter encrustation(CE).Methods Eight patients with difficulty in pulling catheter tube crustcaused were enrolled as our subjects.They were performed pneumatic lithotripsy under ureteroscope operation.The information related to operation was recorded.Results Eight cases were successful in terms of crushing catheter encrustations,and the catheters were removed smoothly.Conclusion The therapy of pneumatic lithotripsy treatment under ureteroscopic is proved to be a minimally invasive treatment with small damage,and higher safety.
6.Effect of conversion of tacrolimus dosage forms from immediate-release to extended-release on kidney function in stable kidney transplant recipients
Xiaohong GUO ; Ning LI ; Mingjun WANG ; Wenping GUO ; Yuan NING ; Ting REN ; Xiaotong WU
Chinese Journal of Organ Transplantation 2021;42(2):104-108
Objective:To observe the effect of conversion from immediate-release tacrolimus (Tac) to extended-release Tac on kidney function in stable kidney transplant recipients.Methods:83 stable kidney transplant recipients who were converted from immediate-release Tac to extended-release Tac in the second people's hospital of Shanxi province from December 2011 to June 2019 were followed up for 12-36 months, and 83 stable kidney transplant recipients who continued to take immediate-release Tac were selected as control group.The changes of kidney function indexes, Tac trough concentration intra-patient variability (IPV) and compliance, the incidence of rejection and the survival rate of grafts and recipients were observed after the conversion from immediate-release Tac to extended-release Tac in stable kidney transplant recipients.Results:The conversion time from immediate-release Tac to extended-release Tac was (42.76±30.50)months after transplantation. At 24 months after conversion, the serum creatinine (SCr) was significantly lower than that before conversion ( P=0.013), and the estimated glomerular filtration rate(eGFR)was significantly higher than that before conversion( P=0.005). In the experimental group , the SCr was significantly lower than that of the control group at 36 months after conversion ( P= 0.017), eGFR was significantly higher than that of the control group ( P=0.038). In the experimental group, the score of Immunosuppressant Therapy Barrier Scale (ITBS) was (20.23±2.89) before conversion and (17.63 ±3.08) after conversion ( P= 0.000). The daily dose of Tac was (2.09 ±0.84) mg before conversion and (2.10 ±0.83) mg after conversion. The trough concentration of Tac before conversion was (7.22 ±2.84) ng/mL, which reduced significantly after conversion. No rejection occurred after conversion, and the recipients/grafts survived healthily during the follow-up period. Conclusions:After conversion from immediate-release Tac to extended-release Tac in stable kidney transplant recipients, the kidney function is stable and better than that of before conversion, the compliance is significantly improved, the IPV of Tac trough concentration is significantly reduced, and long-term use of extended-release Tac has good clinical efficacy and safety.
7.Research on quality parameters of scutellariae radix (formula particles) using on-line NIR in pilot with different extraction process.
Yang LI ; Zhi-Sheng WU ; Xin-Yuan SHI ; Xiao-Ning PAN ; Qiao ZHANG ; Yan-Jiang QIAO
China Journal of Chinese Materia Medica 2014;39(19):3753-3756
The on-line monitor for the changes in the content of baicalin in Scutellariae Radix formula particles during the extraction process was conducted by using near infrared spectroscopy (NIR). High performance liquid chromatography (HPLC) was used as a reference method. Kennard-Stone (KS) was used to divide sample sets, so as to compare different pretreatment methods. The synergy interval partial least squares (SiPLS) was used to screen out modeling wave band to establish partial least-squares models. The relative error method was applied to predict forecast set samples of Scutellariae Radix in three extraction phases. The results showed that the model established by Savitzky-Golay smoothing with 11 points (SG11 points) was the best, with the root mean square with cross validation (RMSECV), root mean square error of correction (RMSEC) and root mean square error of prediction (RMSEP) of baicalin of 0.092 7, 0.134 4 and 0.114 8, respectively, the determination coefficient R2 of greater than 0.99, and the relative error of baicalin content of less than 5%. This indicates that the on-line near infrared reflectance spectroscopy could be applied in on-line monitor and quality control of the extraction process of Scutellariae Radix formula particles.
Automation
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methods
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Drugs, Chinese Herbal
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chemistry
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standards
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Quality Control
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Scutellaria baicalensis
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chemistry
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Spectroscopy, Near-Infrared
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methods
8.Detection analysis of particle size effects in overtone and combination region of NIR based on multi-types of Chinese material medica.
Zhi-sheng WU ; Min DU ; Xiao-ning PAN ; Xin-yuan SHI ; Yan-jiang QIAO
China Journal of Chinese Materia Medica 2015;40(2):287-291
Near-infrared (NIR) diffuse reflectance spectroscopy was used to analyze the impact of multi-class particle size of Chinese material medica (CMM) based on the spectral characteristics in overtone and combination region. Several types of CMM (60, 80, 100,120 mesh) were subjected to NIR spectra analysis. Spectral reproducibility was examined after sample repackage. The result showed that the effects of particle size on the NIR spectra were different according to different bands, in the combination region and first combination-overtone region. Spectroscopy intensity was proportional to the particle size and influence of particle size was greater as the wavelength increased. While in the second combination-overtone region, it was inversely proportional to particle size. To the sampling loading error, the result indicated that when the mesh number was larger than 60 mesh, the error was small. The appropriate particle size was clarified to guarantee the accuracy and reliability of NIR diffuse reflectance spectroscopy in CMM.
Drugs, Chinese Herbal
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analysis
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Materia Medica
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analysis
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Particle Size
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Spectroscopy, Near-Infrared
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methods
9.Rapid discriminant analysis of sulfur fumigated Puerariae Lobatae Radix based on vertical and horizontal surfaces.
Zhi-sheng WU ; Min DU ; Xiao-ning PAN ; Na ZHAO ; Xin-yuan SHI ; Yan-jiang QIAO
China Journal of Chinese Materia Medica 2015;40(12):2336-2339
Near-infrared (NIR) was used as rapid analysis method to identify the sulfur fumigated Puerariae Lobatae Radix. NIR spectra of the cross-sectional and longitudinal selection of samples were acquired. Principal component analysis was conducted. The samples were randomly selected. The different pretreatment methods were compared. Discriminant models were established for every type of spectra to calculate the recognition rate. The orthogonal test and nonparametric test were used to test data normality. The result showed that absorbance values of different sections were different due to the different structure, and the raw spectra were analyzed by PCA method. The result founded that the cumulative contribution rate was arrived at 99.2% while the PC numbers were arrived at 3. The pretreatment method based on the MSC + 1D + Savitzky-Golay was the best to establish the model. For the 50 models constructed with cross-section and longitudinal spectra and total spectra, the recognition rate were (94.4 ± 0.66)%, (94.4 ± 0.66)%, (95.3 ± 0.65)%, respectively, and no difference was observed. The NIR method could be used to identify the sulfur fumigated Puerariae Lobatae Radix.
Chemistry, Pharmaceutical
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methods
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Discriminant Analysis
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Drugs, Chinese Herbal
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chemistry
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Fumigation
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Pueraria
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chemistry
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Spectroscopy, Near-Infrared
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Sulfur
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chemistry
10.Urodynamics and ultrastructure of detrusor in patients with benign prostatic hyperplasia
Jinsheng ZHANG ; Ning LIU ; Yuan QU ; Zongshan WU ; Qian ZHANG ; Jun CHAI ; Chunlin LIU
Chinese Journal of Urology 2008;29(6):422-424
Objective To evaluate the relationship between urodynamics and ultrastructure of bladder detrusor in patients with benign prostatic hyperplasia(BPH). Methods The ultrastructure of detrusor was studied by transmission electron microscopy in 43 cases of BPH(mean age 69.5±6.0 years)and 21 cases(mean age 65.4±7.2 years)of geriatric without bladder outlet obstruction(the controls).All the cases were performed urodynamic test. Results The IPSS(21.1±3.0 VS 7.6±1.4),maximum flow rate(7.7±1.3 ml/s VS 14.9±2.3 ml/s)and AG(44.8±9.9 VS 19.0±5.9)showed significant differences between the the BPH group and control group(P<0.01).Pdet-Qmax in the BPH group(60.1±11.0 cm H20)was higher than that in the control group(48.7±7.1 cm H2o) (P<0.05).The relative density of mitochondria in the BPH group(0.81±0.24)was lower than that in the control group(1.03±0.11)(P<0.05).The intercellular linkage in the BPH group(19.4±4.8 rim)was larger than the control group(14.1±2.0 nm)(P<0.05). Conclusions The swelling and reduction of the relative density of mitochondria could be seen in the bladder detrusor of BPH patients.The large intercellular linkage might be the reason of decrease the coordinate contractility of the blad-der detrusor.