2.Posterior pedicle screw fixation for treatment of fracture and dislocation of atlantoaxial spine
Zexue ZHAO ; Haodong FEI ; Shouguo WANG ; Feng JI ; Yue XUE
Chongqing Medicine 2016;45(10):1350-1352
Objective To investigate the clinical effect of pedicle screw fixation in the treatment of fracture and dislocation of atlantoaxial spine via posterior approach .Methods 19 patients with fracture and dislocation of atlantoaxial spine in this hospital from June 2011 to December 2013 were selected and treated with open reduction and pedicle screw fixation via posterior approach . The X‐radiographs were postoperatively re‐examined at regular time for understanding the correction of fracture and dislocation and implant fusion results ,the neurological functions were evaluated according to the Japanese Orthopaedic Association(JOA) scores . Results All cases got bony fusion without the occurrence of internal fixation loosening ,broken screw or broken rod .The JOA score was improved from preoperative (7 .35 ± 2 .39) points to postoperative (13 .21 ± 2 .53) points (P<0 .05) .Conclusion The posteri‐or atlantoaxial pedicle screw fixation and fusion for treating upper cervical spine injury has satisfactory effect .
3.Construction of Transferring Vector of Marek’s Disease Virus Expressing GFP Gene and Its Primary Application
Ya-Feng QIU ; Fei-Fei GE ; Xue-Lian ZHANG ; Pu-Yan CHEN ;
China Biotechnology 2006;0(05):-
The expressing cassette, LoxP-CMV-gpt-IRES-LoxP( about 2.9kb), was amplified by PCR from a plasmid, pIRES-gpt, by use of the primers , which contained the loxP sites in 5' terminals, respectively. The loxP sites were designed into primers by the software of Primer primer 5.0. Then the cassette was cloned into the site of BalI in pBUS10 to obtain pUS-gptIRES(L). The sequencing analysis for pUS-gptIRES(L) indicated that two loxP sites with the same direction were correctly inserted into pUS-gptIRES(L).The gpt gene in pUS-gptIRES(L) was replaced by a fragment including the full length GFP gene as well as SV40 poly A sequence to get pUS-GFPIRES(L). pUS-GFPIRES(L) was transiently transfected into CHO cell lines, and then the green fluorescence could be seen, the results showed that GFP gene could be expressed correctly. Moreover, pUS-GFPIRES(L) was transfected into the CEF infected MDV CVI988 strain and recombinant virus was selected by the green fluorescence. The growth curve of virus showed the characteristic of recombinant virus was the same as that of CVI988 in vitro. These results give the basis for further studying the characteristic of MDV in vivo and the application of the Cre/LoxP system to MDV genome.
4.Laparoscopic inguinal hernia repair,4 445 cases from a single institution
Fei YUE ; Jianwen LI ; Wenrui WANG ; Ji WANG ; Pei XUE ; Bo FENG ; Minhua ZHENG
Chinese Journal of General Surgery 2016;31(9):724-727
Methods The clinical data of 4445 cases (5 530 hernias) who underwent LIHR at Ruijin Hospital from Jan 2001 to Dec 2015 were analyzed retrospectively.2 125 cases underwent 2 402 trans-abdominal preperitoneal procedure(TAPP),2 306 cases did 2 907 totally extraperitoneal (TEP),and 21 IPOMs in 20 cases.There were 3 216 indirect hernias (60.3%),1 164 direct hernias (21.8%),399 recurrent hernias (7.5%),479 complex hernias (9.0%),and 72 femoral hernias (1.4%).The median time of follow-up is 51 months with a range between 7 and 187 months.Results The average operation time was 27.1 ± 8.7 min for unilateral hernia repair,and 43.0 ± 11.0 min for bilateral hernia repair.The average hospital stay was 1.4 ± 1.1 d.There were 250 seroma (4.7%),68 urinary retention (1.3%),23 transient neuropraxia (0.4%) and 3 paralytic obstruction of intestines (0.1%).Severe complications included 1 port site hernia,1 intestinal injury,and 1 mechanical intestinal obstruction.After a medium follow-up of 51 months,there were 13 recurrent cases (0.24%),including 5 cases after TAPP,7 after TEP,1 after IPOM.Conclusion LIHR is a safe and efficient technique for hernia repair.
6.Research on effect of Sargentodoxae caulis on activity of osteoclasts and proliferation differentiation of osteoblasts.
Li-zhen CHEN ; Ying ZHOU ; Jun-fei HUANG ; Xue ZHANG ; Ting-ting FENG
China Journal of Chinese Materia Medica 2015;40(22):4463-4468
Through morphological observation, HE staining, TRAP staining and toluidine blue staining of bone resorption pits to identify osteoclasts which obtained by 1α, 25-(OH)2 VitD3 inducing rabbit bone marrow cells. Three indicators-TRAP staining, TRAP enzyme activity detecting and the number and area of bone resorption pits were adapted to detect the effect of Sargentodoxae caulis on the activity of osteoclasts. Culturing MC3T3-E1 Subclong 14 cells and detecting the effect of S. caulis on differentiation and proliferation of them by MTT and detecting the alkaline phosphatase in cells. The results show that all of the low, middle and high doses of water and alcohol extracts of S. caulis have significant inhibition on osteoclast differentiation and bone resorption ability in a dose-dependent manner. The low and middle doses of water and alcohol extracts of S. caulis can stimulate differentiation and proliferation of MC3T3-ElSubclone 14 cells, which indicates S. caulis can prevent osteoporosis and the function could be achieved by inhibiting osteoclast activity and promoting the proliferation and differentiation of osteoblasts.
Animals
;
Bone Resorption
;
drug therapy
;
physiopathology
;
Cell Differentiation
;
drug effects
;
Cell Proliferation
;
drug effects
;
Cells, Cultured
;
Drugs, Chinese Herbal
;
pharmacology
;
Humans
;
Mice
;
Osteoclasts
;
cytology
;
drug effects
;
Rabbits
7.Effects of TGF-β stimulation on mouse splenic dendritic cells
Fei YANG ; Ting XUE ; Song CHEN ; Jingru LIU ; Huifang HUANG ; Yuechen LUO ; Fang LIAO ; Xiaoming FENG
Chinese Journal of Microbiology and Immunology 2017;37(3):165-170
Objective To investigate the mechanisms modulating the functions of dendritic cells (DCs) and suppressing the activation and proliferation of T cells by transforming growth factor-β (TGF-β).Methods Mouse splenic DCs were purified with CD11c+ immunomagnetic beads and the purity of isolated DCs were detected by flow cytometry.Gene chip was used to detect gene expression in DCs after stimulation with TGF-β, and then real-time PCR was performed to analyze the differentially expressed genes in microarray at mRNA level.The activation and proliferation of CD4+ T cells which were co-cultured with DCs after stimulation with TGF-β were detected by flow cytometry.Results The purity of DCs reached over 95% after isolation.TGF-β down-regulated the expression of cell surface markers CD53, CD69, CD33, CD74 and CD93 on DCs;decreased the expression of chemokines Ccl3, Ccl5, Ccl9, Ccl6, Ccl17, Cxcl10, Ccl22, Ccl4, Ccr7, Ccl2, Cxcl9 and Ccl7;inhibited the expression of inflammatory cytokines IL-2ra, IL-12rb2, IL-15ra, IL-1b and IL-15.Moreover, the DCs-mediated activation and proliferation of CD4+ T cells were suppressed by TGF-β.Conclusion TGF-β inhibits the DCs-mediated activation and proliferation of CD4+ T cells by suppressing the expression of surface markers on DCs and down-regulating the expression of chemokines and inflammatory cytokines.
8.Clinical efficacy of laparoscopic inguinal hernia repair in elderly patients
Yun ZHANG ; Xiaohui HAO ; Jianwen LI ; Hangjun GONG ; Bo FENG ; Fei LE ; Pei XUE
Chinese Journal of Digestive Surgery 2016;15(10):967-971
Objective To explore the clinical efficacy of laparoscopic inguinal hernia repair (LIHR) in elderly patients.Methods The retrospective cohort study was adopted.The clinical data of 3 203 patients with inguinal hernias (3 847 sides) who were adnitted to the Ruijin Hospital of Shanghai Jiaotong University School of Medicine between January 2001 and December 2013 were collected.Of 3 203 patients,979 (1 107 sides) with age < 60 years and 2 224 (2 740 sides) with age ≥ 60 years were respectively allocated into the under 60 years group and 60 years or older group.The surgical procedures including transabdominal preperitoneal (TAPP) approach,total extraperitoneal (TEP) approach and intraperitoneal onlay mesh (IPOM) approach were selected and performed by doctors in the same team.There were light-weight and heavy-weight patches.Observation indicators included (1) overall operation situations,(2) surgical comparison between the 2 groups,(3)comparison of postoperative indicators between the 2 groups,(4) follow-up.Follow-up using telephone interview and outpatient examination was performed to detect the recovery time of non-restricted activity,recurrence of hernia and complications.Measurement data with normal distribution were represented as ~ ± s and comparison between groups was done by the t test.Comparisons of count data were analyzed using the chi-square test or Fisher exact probability.Ranked data were compared by the nonparametric rank sum test.Results (1) Overall operation situations:3 203 patients with inguinal hernias (3 847 sides) underwent LIHR,including 1 475 (1 677 sides) using TAPP approach,1 718 (2 154 sides) using TEP approach and 10 (16 sides) using IPOM approach (6 using TAPP and IOPM approaches in each side).The light-weight patch was used in 2 206 sides and heavy-weight patch was used in 1 641 sides.Operation time was (31 ± 12) minutes in all 3 203 patients,(27 ±9)minutes in 2 559 patients with unilateral hernia and (44 ± 12)minutes in 644 patients with bilateral hernia,respectively.Duration of postoperative hospital stay was (1.5 ± 1.2) days.(2) Surgical comparison between the 2 groups:TAPP approach,TEP approach,IPOM approach,light-weight patch and heavy-weight patch were performed to 567,538,2,751,356 sides in the under 60 years group and 1 110,1 616,14,1 455,1 285 sides in the 60 years or older group,respectively,with statistically significant differences in above indicators between the 2 groups (X2 =37.976,70.022,P < 0.05).Operation time in unilateral hernia and bilateral hernia and total operation time were (27 ± 9)minutes,(42 ± 10)minutes,(29 ± 10)minutes in the under 60 years group and (27 ± 10)minutes,(44 ± 12)minutes,(3 1 ± 13)minutes in the 60 years or older group,respectively,with no statistically significant difference between the 2 groups (t =-0.106,-1.768,-4.445,P > 0.05).(3) Comparison of postoperative indicators between the 2 groups:the pain score at postoperative day 1 and duration of postoperative hospital stay were 2.4 ± 1.1,(1.5 ± 1.1) days in the under 60 years group and 2.3 ± 1.0,(1.5 ± 1.3) days in the 60 years or older group,respectively,with no statistically significant difference between the 2 groups (t =1.419,-0.126,P >0.05).(4) Follow-up:all the patients were followed up for 23-60 months,with a median time of 43 months.Cases with non-restricted activity recovery at postoperative week 2 and 4 were 973,978 in the under 60 years group and 2 208,2 222 in the 60 years or older group,respectively,showing no statistically significant difference between the 2 groups (X2=0.113,P >0.05).The recurrence of hernia,severe complications,serum tumescence,paresthesia and enteroparalysis were detected in 1,0,49,5,1 sides in the under 60 years group and 11,3,132,16,2 sides in the 60 years or older group,respectively,with no statistically significant difference between the 2 groups (x2=1.556,0.269,0.254,P > 0.05).The urinary retention in the under 60 years group and 60 years or older group was respectively detected in 6 and 44 sides,showing a statistically significant difference between 2 groups (x2=6.956,P < 0.05).Conclusion LIHR is safe and effective in elderly patients,and it can achieve good clinical efficacy under selecting reasonable operation procedures and patches.
9.Antisense oligonucleotides targeting seed sequence of miR-155 and its ap-plications in multiple myeloma
Maoxiao FENG ; Rongxuan ZHU ; Xiaochuang LUO ; Chunming GU ; Xue ZENG ; Jia FEI
Chinese Journal of Pathophysiology 2014;(8):1368-1373
[ABSTRACT]AIM:ToinvestigatetheroleoftinyantisensenucleicacidagainstmiR-155(tinyantimiR-155, t-antimiR-155) in multiple myeloma cells .METHODS:According to the seed sequence of miR-155, t-antimiR-155 was designed and synthesized .t-antimiR-155 was transfected by Lipofectamine TM 2000 into RPMI-8266 cells.The cells were di-vided into t-antimiR-155 group, scrambled control (SCR) group and blank control group .The growth-inhibitory potencies were measured by MTT assay .The ability of cell colony formation was detected by cell colony formation assay .The cell ap-optosis was assessed by flow cytometry with annexin V /PI double staining .RESULTS: The best concentration and time were 0.4 μmol/L and 48 h, respectively.The cell colony forming experiment showed that the circumstances of forming cell community in t-antimiR-155 group was weaker than that in SCR group , and the colony formation inhibitory rate of former was significant higher than the latter .Compared with SCR group , the cell apoptosis in t-antimiR-155 group significantly in-creased.CONCLUSION: The t-antimiR-155 inhibits the progression of multiple myeloma cells by interfering with miR-155.miR-155 may serve as a potential target in gene therapy for treating multiple myeloma .
10.Expression and Identification Truncated Glycoprotein G of Bovine Respiratory Syncytial Virus in Escherichia coli
Jun-Ke FENG ; Fei XUE ; Jiao LI ; Li-Chuang ZU ; Yuan-Mao ZHU ; Xian-Gang REN ;
China Biotechnology 2006;0(12):-
Two fragments G1 and G2 of the glycoprotein G gene of bovine respiratory syncytial virus(BRSV) were selected for expression in Escherichia coli based on the analysis of glycoprotein G by DNA Star software.Then the two fragments of glycoprotein G were amplified by PCR with synthesized G gene of BRSV as the template.The amplified fragments G1 and G2 are 570bp and 308bp in length,respectively.The PCR products were cloned into pET30a vector and expressed in soluble form in E.coli after induction of cultured E.coli with IPTG.Both of the recombinant proteins G1 and G2 were purified by immobilized Ni ion affinity chromatography under native conditions.Then the purified proteins were analysed by Western blotting.The results showed that the purified recombinant protein G1 retained good antigenicity and specificity.But the purified recombinant protein G2 didn't possess biological activity.Antibodies against BRSV were detected in suspected bovine serum samples in China by using indirect ELISA and Western blotting with the purified recombinant protein G1.The purified recombinant protein G1 might be used as antigen for establishing serological methods for diagnosis of BRSV infection.And the purified recombinant protein G1 might also be used for preparing polyclonal and monoclonal antibodies for research on biological functions of glycoprotein G of BRSV.