2.Construction,expression of I-Ad/IgG2b Fc dimer fusion protein and its identification
Zhihuan LUO ; Hang QIAN ; Chengwei XU ; Xinwen MIN ; Jun CHEN
Chinese Journal of Immunology 2017;33(4):498-501,506
Objective:To construct I-Ad/IgG2b Fc baculovirus expression vector and express I-Ad/IgG2b Fc dimer fusion protein in Sf9 insect cells.Methods:I-Ad α,I-Ad β and IgG2b Fc gene sequences were amplified from BALB/c mouse lymphocytes by RT-PCR.I-Ad α and I-Ad β were connected with the leucine zipper sequence Fos and Jun respectively by overlapping PCR to form I-Ad α-Fos and I-Ad β-Jun.I-Ad α-Fos and IgG2b Fc fragments were ligated by restriction sites Xba I to form I-Ad α-Fos-IgG2b Fc recombination sequence.I-Ad α-Fos-IgG2b Fc and I-Ad β-Jun fragments were inserted to PPH and PP10,which were the downstream of the promoters in the plasmid pFastBacTMDual,to form pFastBacTMDual+[I-Ad/IgG2b Fc] recombinant plasmids.The constructed vector was identified by PCR,restriction endonuclease and sequencing.The recombinant plasmids pFastBacTMDual+[I-Ad/IgG2b Fc] was transferred into the DH10Bac competent cell to form recombinant baculovirus Bacmid+[I-Ad/IgG2b Fc].The recombinant baculovirus was transfected into Sf9 insect cells by liposome transfection reagent.After infected with Sf9 insect cells,the supernatant was collected and concentrated by PEG20000 to obtain I-Ad/IgG2b Fc dimer fusion protein.The fusion protein was detected by double-antibody sandwich ELISA and Western blot.Results:PCR,restriction enzyme digestion and sequencing confirmed that the recombinant vector pFastBacTMDual+[I-Ad/IgG2b Fc] had the correct sequence.The double antibody sandwich ELISA and Western blot showed that recombinant bacmid could successfully infect Sf9 insect cells,and the expressed fusion protein had the correct conformation.Conclusion:The pFastBacTMDual+[I-Ad/IgG2b Fc] baculovirus expression vector was successfully constructed and expressed in Sf9 insect cells,laying a foundation for the study of I-Ad-restricted T cells.
3.Survey on anxiety-depression emotion on 100 tumor patients
Juan PENG ; Lujiao ZHOU ; Hang ZHOU ; Qian YIN
Chongqing Medicine 2015;(14):1949-1950,1953
Objective To investigate and analyze the anxiety‐depression emotion in tumor patients in a tumor hospital to pro‐vide the scientific basis for mental intervention of the tumor patients .Methods Totally 100 tumor patients were surveyed by using the Beck Depression Inventory(Beck) ,Self‐Assessment Anxiety Scale(SAS) and self‐designed general condition information ques‐tionnaire .Results Among 100 tumor patients ,86 cases (86% ) had the depression emotion and 90 cases(90% ) had the anxiety e‐motion .In the comparison of anxiety‐depression emotion among different tumor sites ,disease entities ,genders ,age and educational levels ,except the depression emotion had the statistical difference between the patients with primary school degree and the patients with the cultural level higher than junior middle school (P<0 .05) ,the comparison in other indexes had no statistical differences (P>0 .05) .Conclusion The patients with tumor had higher occurrence rate of anxiety‐depression emotion .The mental intervention should be paid attention to improve their life quality .
4.Change of Serum Neuroglobin in Newborn Infants with Fetal Distress
jing, WANG ; hang-yan, WANG ; ping, WANG ; qian, LI
Journal of Applied Clinical Pediatrics 1986;0(02):-
Objective To observe the change of serum neuroglobin(NGB)of normal full-term and fetal distress infants,and to explore the sensitivity and validity of NGB as potential biomarker for brain injury.Methods The technique of double-antibody sandwich enzyme-linked immunosorbent assay to measure NGB was established,and stable data standard curve was gained.Serum NGB of umbilical cord artery in 35 normal full-term(17 male,18 female)and 32 newborn infants were measured.The differece of serum NGB in gender,and the changes in serum NGB of fetal distress infants were analyzed,who were probably injured by anoxia.The results were analyzed by Stata 7.0 software.Results The standard curvilinear equation was Y=exp(-5.881 532+11.955 890?X)+2.281 506E-02(linear correlation coefficient r=0.999 4 P0.05);the average NGB in the serum of umbilical cord artery of fetal distress infants was(149.7?43.2)?g/L,which was significantly higher than that of normal full-term infants(t=2.941 6 P
5.Retrospective analysis of perioperative complications and risk factors of posterior lumbar interbody fusion for recurrent lumbar disc herniation
Xinhua ZHAO ; Hang YUAN ; Jinqian QIAN ; Yonghong YANG
Chinese Journal of Orthopaedics 2016;36(17):1121-1125
Objective To retrospectively analyze the perioperative complications of posterior lumbar interbody fusion (PLIF) for recurrent lumbar disc herniation and identify potential risk factors that correlate with those complications.Methods All of 71 patients with recurrent lumbar disc herniation were treated surgically with PLIF,discharged from our department between January 2008 and December 2014.Demographic and operation data were collected and perioperative complications were recorded.We analyzed whether the clinical factors (age,gender,BMI,co-morbidity,smoking,time of recurrence,blood loss,operation segment,operation time) were in correlation with perioperative complication by univariate analysis.Then we integrated the statistically significant indicators into Logistic regression equation to determine the related risk factors for complication.Results The study group consisted of 71 cases,including 42 males and 29 females.The age was 19 to 64 years old with an average of 50.6 years old,and the average BMI was 23.6 kg/m2.26 cases had perioperative complications,while there were two or more complications in 5 patients,and no mortalities.Neurologic deterioration or neuropathic pain (10 cases,14.1%) and dural tears (6 cases,8.5%) were the most common intraoperaitive complications.The other complications included nerve root or cauda equine injury,deep or superficial wound infection,urinary tract infection,respiratory system complication,cardio-vascular complication and delirium.Univariate analysis suggested that age,gender,co-morbidity,smoking,time of recurrence,operation segment,operation time were not associated with perioperative complication.The multivariate Logistic regression analysis showed BMI and blood loss were closely related to perioperative complication.Conclusion Complications of posterior spinal fusion surgery for recurrent lumbar disc herniation are affected by many factors.The most common complications are transient neurologic deterioration or neuropathic pain and dural tears.BMI and blood loss are independent risk factors.
6.The relationship between proteolysis-inducing factor and the nutritional status and prognosis of gastric cancer patients
Yuanshui SUN ; Fengyong WANG ; Hang LU ; Xiaoyu QIAN
Journal of Chinese Physician 2015;17(5):644-647
Objective To investigate the prognostic significance of proteolysis-inducing factor (PIF) in gastric cancer patients.Methods Biopsies of the rectus abdominis muscle were obtained intra operatively from 53 gastric cancer patients and 53 subjects undergoing surgery for benign abdominal diseases,muscle and serum PIF,ubiquitin expression,and proteasome proteolytic activities were assessed through reverse-transcription polymerase chain reaction (RT-PCR),Western blot,and enzyme-linked immunosorbent assay (ELISA).Results PIF was significantly upregulated in muscle of gastric cancer compared to control muscles.PIF was upregulated in 60% (32/53) muscle of gastric cancer.Over expression of PIF in muscles of gastric cancer were associated with level of serum albumin,tumor node metastasis (TNM) stage,and percent of weight loss.The survival rate of patients with a high expression of PIF was significantly lower than those in patients with low expression (P < 0.05).Conclusions PIF was significantly upregulated in muscle of gastric cancer,over expression of PIF in muscles of gastric cancer was associated with level of serum albumin,TNM stage,percent of weight loss,and prognosis.PIF might play an important role in progression and prognosis of gastric cancer.
7.The Potential Alteration of Their Osteogenesis and Adipocyte Differentiation in Bone Marrow Mesenchymal Stem Cells from Mice with Immune-mediated Aplastic Anemia
Ruilan GAO ; Liming YIN ; Xudai QIAN ; Zhou HANG
Journal of Zhejiang Chinese Medical University 2006;0(05):-
[Objective]To explore the potential alteration of their osteogenesis and adipocyte differentiation in bone marrow mesenchymal stems cells from mice with immune-mediated aplastic anemia.[Methods]Balb/c mice model of immune-mediated aplastic anemia was established by radiation with sublethal dose of 60Co following the intravenously infusion lymphocytes of DAB/2 mice.The culture of the MSC and CFU-F and pathological examination of bone marrow were carried out 15 days later.The amount of calcium node and the frequency of adipocyte differentiation were evaluated respectively by alizarin red and oil red O.[Result]The number of CFU-F and the number of calcium node in model mice decreased more greatly than normal mice,but the ferenqucy of adipocyte differentiation was more increased greatly in model mice than normal mice;the pathological examination showed in the model mice,the hematopoietic structure was destroyed and filled with abundant adipocyte.[Conclusion]The potention of osteogenesis and adipocyte differentiation was altered in the mice with immune-mediated aplastic anemia.
8.Development of A/B Whole-automatic Machines for Dispensing, Filtering and Filling the Drug Solution
Linkuan WANG ; Wengyu SHI ; Yun CHEN ; Baoming HANG ; Yongsheng HU ; Cunsheng QIAN
Chinese Medical Equipment Journal 2003;0(10):-
Objective To develop an automatic machine used for dispensing, filtering and filling the drug solution. Methods The microcomputer integrated with photoelectric -machinery technique is used to develop the automaitically opertating machines which have series of automatic functions,such as heating, stirring, dispensing, filtering and filling the drug solution, as well as digitally dispensing amount, heating temperature, filling bottle-number, and etc. Results The machine is suitable for automatically preparing,filtering a volume of 5,000ml to 30,000ml and automatically filling various specifications of drug solution with a volume of 30ml to ≥1,000ml in all kinds of army and locality hospital pharmacy. Conclusion This machine is characterized with novel and airtight design, and complete functions. It is suitable to be used not only in common but also under field condition of army.
9.Prognostic value of pre-chemotherapy serum cystatin C in patients with castration-resistant prostate cancer
Lulu FAN ; Chenggong LIAO ; Jianguo HUANG ; Hang YIN ; Menlong QIAN ; Nao WAN ; Ning LU
Journal of International Oncology 2017;44(5):356-360
Objective To explore the prognostic value of pre-chemotherapy serum cystatin C (Cys C) in patients with castration-resistant prostate cancer (CRPC).Methods The medical records of 48 patients with CRPC were reviewed.These patients were diagnosed and underwent docetaxel-based chemotherapy in Xinjiang Military Command General Hospital between January 2009 and January 2015.Statistical analysis was performed to identify the clinical and prognosis value of Cys C.Results Of 48 patients with CRPC,the expression of serum Cys C before chemotherapy was related with Gleason score (x2 =4.218,P =0.040) and distant metastasis(x2 =4.090,P =0.043).23 patients was in high group (Cys C > 1.61 mg/L),25 in low (Cys C ≤ 1.61 mg/L).The median survival time of high group and low group were 15.6 and 25.3 months,respectively (x2 =13.876,P < 0.001).Univariate analysis showed that TNM stage (x2 =6.934,P =0.018),Gleason score (x2 =7.933,P =0.005),baseline prostate specific antigen (PSA) (x2 =9.038,P =0.003),number of chemotherapy cycles (x2 =5.024,P =0.028),distant metastases (x2 =6.963,P =0.013) and serum Cys C before chemotherapy (x2 =6.976,P =0.012) were associated with overall survival of patients with CRPC.COX multivariate analysis showed that baseline PSA at diagnosis (x2 =4.257,P =0.039),number of chemotherapy cycles (x2 =6.245,P =0.017),distant metastases (x2 =5.122,P =0.028) and serum Cys C before chemotherapy (x2 =8.172,P =0.004) were independent risk factors of overall survival of patients with CRPC,especially serum Cys C before chemotherapy (HR =2.394).Conclusion The patients with high Cys C level have poor prognosis,and the pre-chemotherapy Cys C is an independent risk factor for prognosis of CRPC patients treated with docetaxel-based chemotherapy which can be used as an effective indicator to assess the prognosis of CRPC.
10.Expression and Clinical Significance of Bcl-6 mRNA in the T Follicular Helper(Tfh)Cells of HIV/AIDS Patients
Qian FENG ; Weihong REN ; Yanqing LI ; Wenbo LI ; Yan TONG ; Hang ZHAO ; Huijie FENG
Journal of Modern Laboratory Medicine 2015;(3):64-66,70
Objective To explore the expression of Bcl-6 mRNA in the Tfh cells of HIV/AIDS patients and the relationship between Bcl-6 mRNA and the progression of HIV/AIDS.Methods This experiment chose 60 patients who were confirmed by HIV antibody test positive,during May 2014 to November from AIDS Research Institute in the First Affiliated Hospi-tal,Henan College of Traditional Chinese Medicine.According to the amount of CD4+T cells,the patients with HIV/AIDS were divided into A(CD4<200 cells/μl),B(200 cells/μl