3.Prokaryotic expression and purification of human Smith D1 antigen
Wen-Bing WU ; Xiao-Peng LAN ; Xiang-Yue YANG ;
Chinese Journal of Laboratory Medicine 2000;0(06):-
Objective To obtain recombinant human Smith D1 (Sm D1) antigen and establish detecting assay.Methods Human Smith D1 antigen was synthesized by PCR using human Leukemic cDNA. The prokaryotic expression vector pGEX-ST-Sm D1 was constructed and transformed into E.coli.BL21 cell.Protein expressed under the induction of IPTG.We established DIGFA for detecting anti-Sm D1 antibodies with purified Sm D1 antigens.Results Sequence and restriction analysis revealed Sm D1 gene was cloned in frame into pGEX-5T,SDS-PAGE profile showed a clear protein band with a relative molecular weight of 39 000 and western blotting indicated that the expressed product specifically reacted to polyclonal anti-human Sm D1 genes.There was no significant difference between DIGFA and IB.The agreement between DIGFA and IB was 91.7% as calculated by Kappa statistical method.The sensitivity and specificity of DIGFA were 100% and 83.3% repectively.Conclusions Human Sm D1 gene is successfully cloned、 expressed and purification.The DIGFA,using purified Sm D1 antigens,is as good as IB,rather simpler, more rapid and reliable assay.
4.Eukaryotic Expression and Primarily Application of Human Smith D1 Antigen in Methylotrophic Yeast Pichia pastoris
Xiang-Yue YANG ; Wen-Bing WU ; Xiao-Peng LAN ;
Microbiology 1992;0(01):-
To clone, express and primarily use human autoantigen Sm D1 in methylotrophic yeast Pichia Pastoris. The gene Sm D1 was cloned by PCR.The PCR product was inserted into the vector pPIC9k. The recombinant plasmid pPIC9k- Sm D1 was transformed into yeast SMD1168 by electroporation. The positive clones were screened in MD plates. The high copy number transformants were rapidly selected by using G418 and were induced by methanol. Supernatants after induction were analyzed by SDS-PAGE and im-munodot. The PCR product was showed about 360 bp in size which was in accordance with predicted. The pPIC9k-Sm D1 showed the same seqencing result with GenBank’s report and restriction enzyme analysis confirmed our prediction. The pPIC9k-Sm D1 positive clone produced an about 16 kD protein which had natural immunogenicity of human autoantigen Sm D1 by SDS-PAGE and immunodot. The sensitivity and specificity of immunodot were 96% and 100%, respectively. The agreement between immunodot and im-munoblot was 98%. Successfully cloning and high-level expression of human autoantigen Sm D1 in methy-lotrophic yeast Pichia pastoris laid a foundation for further research work.
5.Evaluation of clinical effect of porcelain fused for metal crown restoration after root canal therapy
Hong WANG ; yu Xiang WANG ; Wei REN ; bing Xiang WU
Chinese Medical Equipment Journal 2017;38(8):72-74,85
Objective To investigate the clinical effect of porcelain fused for metal crown restoration after root canal therapy.Methods Totally 90 patients with pulpitis or periapical periodontitis from December 2013 to December 2015 had their clinical data analyzed,who underwent root canal therapy and were divided equally into an observation group and a control group.The observation group took porcelain endocrowns repair and the control group applied ceramic crown.The two groups were compared on the restoration integrity,edge sealing,residual rate,gum,food impaction and color 3 months,6 months and 1 a after treatment.Results The observation group behaved significantly better than the control group in the edge sealing and restoration integrity (P<0.05).There were significant differences between the residual rates,gum and food impaction in the two groups (P<0.05).Conclusion Porcelain endocrowns repair gains advantages over ceramic crown repair in edge sealing,integrity,gum,food impaction and color after root canal therapy,though its long-term effect needs further investigation.
7.Entity of acupoint: kinetic changes of acupoints in histocytochemistry.
Wei HE ; Meiling WU ; Xiang-hong JING ; Wanzhu BAI ; Bing ZHU ; Xiaochun YU
Chinese Acupuncture & Moxibustion 2015;35(11):1181-1186
The present study aims to investigate the kinetic histocytochemical changes of acupoints in different condition. The expression of tryptase (+) mast cells, histamine (HA) , serotonin (5-HT) and nociceptive neuropeptides including calcitonin gene-related peptide (CGRP) and substance P (SP) were observed by immunohistochemistry combined with confocal technology. Mast cells were labeled with anti-mast cell tryptase antibody and simultaneously with HA or 5-HT primary antibodies to observe their co-expression. The results showed that: (1) SP and CGRP were expressed more highly on the cutaneous nerve fibers of "Hegu" (LI 4) after acupuncture stimulation than that of the control. Mast cells aggregated in close proximity to the blood vessels in intra-epidermis and dermis, and some of them with degranulation in the lower dermis and subcutaneous tissue of "Hegu" (LI 4). Both mast cells and their granules appeared with HA (+) and 5-HT (+) expression at stimulated LI 4 sites, while a few intact mast cells with a little expression of 5-HT and HA were distributed in areas of non-stimulated Ll 4. (2) The acupoints in different locations such as Baihui (GV 20), Weishu (BL 21), Zhongwan (CV 12) and LI 4 had the same constituent but the contents were different. (3) The histocytochemical responses of acupoints sensitized by the Gastric mucosa injury (GMI) were also investigated. GMI resulted in neurogenic plasma extravasation by Evans Blue (EB) in the skin of the acupoints over the back and abdomen, which mostly occurred in the T9-T11 dermatomere. The EB extravasation dots just like acupoints sensitization appeared after GMI and disappeared gradually during the natural self-recovery of the gastric mucosa. More SP and CGRP positive nerve fibers were distributed in EB dots than in regions beside EB dots and in the control, mostly distributed in the nerve fibers around both the vessels and root of hair follicle. Mast cells also aggregated and degranulated to release algogenic substances of 5-HT and HA around the vessels in areas of the EB dots. Collectively the acupoints displayed the same histocytochemical responses due to either acupuncture stimulation or GMI. This may potentially be the histocytochemical basis in the local acupoints and acupoints displayed kinetic changes in different condition.
Acupuncture Points
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Acupuncture Therapy
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Animals
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Calcitonin Gene-Related Peptide
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metabolism
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Gastric Mucosa
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chemistry
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metabolism
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Histocytochemistry
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Humans
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Mast Cells
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chemistry
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metabolism
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Mice
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Rats
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Serotonin
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metabolism
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Skin
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chemistry
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metabolism
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Substance P
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metabolism
8.Endoscopic unilateral versus bilateral biliary stenting for the palliation of malignant hilar obstruction:a meta-analysis
Yubao ZHOU ; Jun WU ; Daoquan ZHANG ; Shu JIN ; Xiang ZHAN ; Bing HU
Chinese Journal of Digestive Endoscopy 2016;(1):49-53
Objective To evaluate the efficacy and safety of the unilateral and bilateral drainage in hilar malignant biliary obstruction with a meta?analysis of relevant studies. Methods A systematic electronic search with keywords “ biliary stent”, “hilar tumor” and “malignant biliary obstruction” was independently performed by two reviewers in major electronic databases of Pubmed, EMBASE, Cochrane Library and Web of Science. A meta?analysis was conducted using Revman 5?? 1 software. Results Six studies were included with a total number of 558 patients, of whom 307 patients underwent unilateral biliary drainage and 251 pa?tients bilateral drainage. There was no significant difference in early complications ( OR = 1?? 03,95% CI:0?? 58?1?? 81,P = 0?? 93), later complications (OR = 0?? 98,95%CI:0?? 46?2?? 07,P = 0?? 95), operation success rate (OR= 1?? 42,95%CI:0?? 76?2?? 66,P = 0?? 28),drainage efficacy or median survival time between the two groups. Compared with unilateral biliary drainage, bilateral biliary drainage had a longer time of stent patency (MD = - 29?? 12,95% CI:- 38?? 55?- 19?? 70,P< 0?? 001). Conclusion Both unilateral and bilateral biliary drainage are safe and effective for hilar malignant biliary obstruction, but more high quality clinical researches are needed.
9.Effects of serum estradiol levels during controlled ovarian hyperstimulation on outcomes of IVF-ET
Xiang, LU ; Lu, LI ; Xiao-hong, GAO ; Yu, WU ; Bing, XU ; Xiao-xi, SUN
Journal of Shanghai Jiaotong University(Medical Science) 2009;29(7):850-853
Objective To analyse the effects of serum estradiol levels during controlled ovarian hyperstimulation (COH) on the outcomes of in vitro fertilization and embryo transfer (IVF-ET). Methods The clinical data of 472 patients undergoing IVF-ET with GnRH analogues recombinant FSH long protocol were retrospectively analysed. The area under the curve (AUC) of estradiol (E2) level was calculated during COH, and patients were categorized into groups according to the percentile of AUC of E2(AUCE2) during COH. The general characteristics and parameters related to the outcomes of IVF-ET were compared among groups. Results The 10th percentile and 90th percentile of AUCE2 were 3 347.0 pmoL/L and 14 414.3 pmol/L, respectively. Four hundred and seventy-two patients were divided into lower reaction group (AUCE2 3 347.0 pmol/L, n=48), normal reaction group (14 414.3 pmol/L>AUCE2 > 3 347.0 pmol/L, n=376) and higher reaction group (AUCE2≥14 413.3 pmol/L, n=48). There was no significant difference in age, body mass index, baseline follicle stimulating hormone level, time of treatment with gonadotropin, endometrium thickness on day of transfer and embryos transferred(P>0.05). Compared with lower reaction group and normal reaction group, the number of oocytes per retrieval and number of embryos frozen were significantly larger(P<0.01) and the mild/severe ovarian hyperstimulation syndrome rate was significantly higher in higher reaction group(P<0.05). There was no significant difference in fertilization rate, cumulative embryo score, high-grade embryo rate, clinical pregnancy rate and implantation rate among groups (P> 0.05). Conclusion Sustained snpraphysiological serum E2 levels during the COH process do not adversely affect the quality of oocytes and embryos, clinical pregnancy rate and implantation rate to some extent in IVF-ET.
10.Therapeutic effects of titanium elastic nails for the treatment of clavicular fractures in elder children.
Guo-jun XU ; Guo-lin WU ; Xu-hong CHEN ; Xiang-rong JI ; Hong-bing HUANG ; Chao-lin CHEN
China Journal of Orthopaedics and Traumatology 2015;28(2):101-105
OBJECTIVETo research the application and clinical effect of titanium elastic nails (TEN) for the treatment of closed clavicular fracture in elder children.
METHODSFrom October 2010 to December 2012,16 elderly children with clavicular fracture of elder children were treated with internal fixation, including 9 boys and 7 girls with an average age of 14.2 yeats old ranging from 9 to 17 years. The mean time between injury and surgery was 2 to 6 days. Thirteen patients had a transverse fractures; 3 patients had a oblique fractures, excluding the long-oblique patients. Neer function score after 3 months follow-up and the shoulder activity degree after 3 weeks followed were compared respectively between the injured side and healthy side. Fracture reduction and healing were followed up by X-rays to analyze internal fixation with TEN technique.
RESULTSAll patients were healed well after surgery. There were 2 cases with soft tissue irritation. No cases of infection, TEN broken, delayed healing or skin bursting were observed. All patients were followed up for 3 to 10 months,with an average of 7.2 months. There were no significant differences (t=2.12, P>0.05) between the healthy side (98.75±2.70) and injured side(96.69±6.22)of Neer score at 3 months during followed up. The shoulder activity at 3 weeks during followed up was different between both sides (P<0.05). The intramedullary nail was removed after X-rays at 12-18 weeks during followed up showed good bone healing,and the shoulder function was recovered well.
CONCLUSIONInternal fixation with TEN for the treatment of clavicular fracture has advantages of a limited invasion, beauty, safety and reliability. This technique provides an ideal fixation option for the treatment of clavicular fracture in elder children.
Adolescent ; Bone Nails ; Child ; Clavicle ; injuries ; surgery ; Elasticity ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Titanium