1.Prokaryotic expression,purification and polyclonal antibody preparation of C terminal Helicase domain of mouse Rig-I
Jinju LI ; Hua REN ; Ziqiang WANG ; Min QIAN ; Bing DU
Chinese Journal of Immunology 2000;0(09):-
Objective:To develop a highly efficacious and sensitive immunological reagent for further investigation on the retinoic acid-induced gene I (Rig-I) of mouse .Methods:The Helicase domain coding region (726-2 240 bp) of mRig-I-H was cloned into plasmid pET15b (+) to construct the recombinant plasmid pET15b(+)-mRig-I-H.Then the plasmid was transformed into E.coli BL21 for protein expression.Rabbits were immunized with electrophoresis-purified recombinant protein to obtain the polyclonal antibody against mRig-I-H.The titer of polyclonal antibody was detected by ELISA and the specificity was identified by Western blot and Immunofluorescence.Results:The recombinant protein was expressed successfully in E.coli.Western blot analysis showed that target protein was expressed with a molecular weight of 40 kD.Titer of the polyclonal antibody was about 1∶1?105 by ELISA assay.With this antibody,we could detect the expression of Rig-I in RAW 264.7 cell line by Western blot and Immunofluorescence.Conclusion:The high level expression of Rig-I Helicase domain is induced in E.coli expressing system.Anti-mRig-I-H polyclonal antibody with high titer and fine specificity could be a novel tool in future investigation of Rig-I.
2.Perioperative nutrition management of liver transplantation patients
Wei LU ; Shi FANG ; Shuyu ZHUO ; Yanbin YE ; Ziqiang TAI ; Shikun QIAN
Parenteral & Enteral Nutrition 1997;0(04):-
Objective:To summarize the perioperative nutrition management,improve the successful rate in liver transplantation patients and reduce the complications after operation.Methods:A retrospective analysis of the perioperative nutrition treatment was made in 71 cases with liver transplantation.The principle of enteral nutrition before operation was appropriate energy,protein and rich vitamin,and the branched-chain amino acid(BCAA) and glutamine(Gln) were supplied.Intravenous nutrition,early enteral nutrition and total enteral nutrition support were applied during the postoperation period.Results: Except 5 cases that died from respiratory failure and 2 died from hemorrhagic shock respectively,other 64 cases recovered gradually in liver function,and their nutrition indices were enhanced significantly.Conclusion:The perioperative nutrition management is very necessary for liver transplantation patients,and the appropriate nutrition support is helpful to rehabilitate the function of graft organ and the nutritional status of the recipients as possible as early.
3.Repeated computed tomography scanning in assessing the change of tumor bed volume during whole breast irradiation in early-stage breast cancer after breast conservative surgery
Zhaozhi YANG ; Gang CAI ; Ziqiang PAN ; Jiayi CHEN ; Xiaomao GUO ; Xiaoli YU ; Qian ZHANG ; Xin MEI ; Jiongyan LI
Chinese Journal of Radiation Oncology 2010;19(6):524-527
Objective To determine the change of tumor bed volume during whole breast irradiation by repeated computed tomography scanning and to analyze the dosimetric impact of boost-planning on different CT images. Methods From July 2008 to Jan 2009, sixteen patients with early-stage breast cancer underwent breast conservative surgery (BCS) were enrolled in the study. All patients received whole breast irradiation and tumor bed boost, no adjuvant chemotherapy was given. Two additional CT scans were acquired in addition to the planning CT ( CT1 ), one in the course of radiotherapy ( CT2 ) and the other before the boost (CT3). Tumor beds were contoured in all CT images. Three-dimensional conformal radiotherapy planning for tumor bed boost was done on CT1 and CT3 respectively. Results The mean tumor bed volume on CT1, CT2 and CT3 were 49.5 cm3, 25.6 cm3 and 22. 2 cm3 ( F = 5. 63, P = 0. 007 ),respectively. Further analysis found statistically significant difference between CT1 and CT2 ( q = 0. 03, P =0. 010), CT1 and CT3 ( q = 0. 01, P = 0. 004), but not between CT2 and CT3 ( q = 1.00, P = 0. 333 ). The average reduction of tumor bed volume from CT1 to CT3 was 43.4%. A reduction of 20% or above was found in 88% of the patients ( n = 14), 50% or above in 38% of the patients (n = 6). In the boost-planning, the volume of the ipsilateral breast receiving 100% prescribed dose (V100%) on CT1 and CT3 was 183.5 cm3 and 144. 5 cm3, respectively ( t = 3.06, P = 0. 008 ). Conclusions Volume of tumor bed is dynamically reduced in the course of whole breast irradiation after BCS, with more important reduction in the early weeks after the beginning of irradiation. A second CT scan before tumor bed boost is warranted.
4.Clinical analysis of 11 cases of urothelial carcinoma in the ipsilateral kidney transplant recipients
Zhanpo YANG ; Wenhui ZHOU ; Ziqiang XU ; Qian LIU ; Hongshun MA
Chinese Journal of Organ Transplantation 2017;38(9):555-558
Objective To explore the clinical characteristic,treatment,and prognosis of urological de novo malignant tumor in kidney transplant recipients.Methods A retrospective analysis was performed on 11 patients with urothelial carcinoma admitted in our institute between 2008 and 2016.Three patients were male and 8 patients were female.The interval between tumorigenesis and transplantation ranged from 12 to 132 months with a mean time of 68.4 months.Of the 11 cases,5 had pelvic TCC,4 cases had ureter TCC,and the rest 2 cases had pelvic and ureter TCC.All patients were in the same side of transplanted kidney (right).Of the 11 cases,7 had a main clinical manifestation of gross hematuria,3 cases had abdomen pain of the right side,and 1 case had hydronephosis of the right side during physical examination.Surgical treatment was taken in all 11 cases,combined with chemotherapy and irnmunotherapy (decreased immunosuppressive agents)treatment.Results Surgical treatment in all 11 cases was successful,and the postoperative pathology results confirmed the diagnosis of urothelial carcinoma.One patient receiving palliative treatment died 4 months after diagnosis.One patient died of extensive metastatic disease at 36th month postoperatively,and one patient died of respiratory failure.The rest 8 cases were followed for 4-96 months.One patient was given sirolimus (SRL) but diarrhea,so cyclosporine A (CsA) was administered.The renal function in the remaining 8 patients was normal.Conclusion Laparoscopic surgery combined with middle and small incision in the treatment of autologous ipsilateral urothelial carcinoma after renal transplantation is safe and effective.
5.Vaginal microbiota characteristics and influencing factors in normal pregnant women
Yaxin LI ; Zongguang LI ; Ziqiang QIAN ; Miao ZHANG ; Hui KAN ; Yutong MU ; Yanmin CAO ; Yao DONG ; Kailin WANG ; Yijie LI ; Haiyan LIU ; Qing LI ; Anqun HU ; Yingjie ZHENG
Chinese Journal of Microbiology and Immunology 2022;42(1):50-61
Objective:To study the characteristics and influencing factors of vaginal microbiota in normal pregnant women.Methods:This study was based on a cohort of pregnant women established in Anqing Municipal Hospital Affiliated to Anhui Medical University from February 2018 to February 2020. Vaginal samples of normal pregnant women who met the inclusion and exclusion criteria were ordered by the gestational weeks at sampling. Five samples were randomly selected from each gestational week group and if the samples were less than five, all samples were included. Sequencing of the V3-V4 region of the 16S rRNA gene was performed. Dominant species were analyzed by MicrobiomeAnalyst. Alpha diversity was measured with Chao1, Observed Features, Shannon diversity, Simpson diversity, Faith_pd and Pielou′s Evenness. The dominant status of Lactobacillus was also described and compared. Multiple linear regression and logistic regression were used to analyze the factors influencing vaginal microbiota. Analysis of variance and Kruskal Wallis test were used for statistical analysis of continuous variables, and Chi-square test and Fisher′s exact test were used for categorical data. The differences were considered statistically significant when the P value was less than 0.05. Results:This study enrolled 91 pregnant women (91 vaginal samples) with an average age of (27.37±3.60) years. There were 18, 56 and 17 vaginal samples collected at the median gestational age of 11.93 weeks (the first trimester), 19.43 weeks (the second trimester) and 38.29 weeks (the third trimester), respectively. The relative abundance of Firmicutes and Lactobacillus was 91.30% and 87.67%, respectively. Lactobacillus iners and Lactobacillus crispatus had a relative abundance of 43.95% and 36.33%, respectively. Moreover, Lactobacillus iners-dominated vaginal microbiota was detected in all trimesters. The number of samples with high relative abundance of Lactobacillus iners gradually decreased with gestational age. Lactobacillus crispatus-dominated vaginal microbiota was found in the second and third trimesters and the number of samples with high relative abundance gradually increased during pregnancy. The Alpha diversity of vaginal microbiota had a decreasing trend during the gestation. There were significant differences in Pielou′s Evenness diversity index of vaginal microbiota between different smoking groups ( P<0.05) and in Shannon diversity index between different drinking groups ( P<0.05). There were significant differences in Chao1, Observed Features and Faith_pd diversity index of vaginal microbiota between pregnant women with different education ( P<0.05) and in Shannon and Simpson diversity index between different income groups ( P<0.05). Conclusions:Vaginal microbiota was dominated by Lactobacillus in normal pregnant women. The dominance of Lactobacillus iners gradually decreased, while that of Lactobacillus crispatus increased during gestation. In normal pregnant women, the Alpha diversity of vaginal microbiota was correlated with smoking, drinking, education and family annual income. Smoking cessation and drinking before pregnancy were related to lower Alpha diversity of vaginal microbiota in pregnant women, while lower education and higher family income were associated with higher Alpha diversity.