1.The immunological and clinical significance of IgM in primary biliary cirrhosis
Chinese Journal of Laboratory Medicine 2013;(3):201-205
Primary biliary cirrhosis (PBC) is a slowly progressive autoimmune disease of the liver.Studies on the pathphysiology of this disease have demonstrated the immune-mediated destruction of the intrahepatic bile ducts and portal infammation seem to be the result of an intense autoimmune response directed against the autoantigens located in the biliary epithelium.Immunologically,PBC is characterized by the presense of antimitochodrial antibodies,elevated serum IgM and lymphoid infiltration in the portal tract.Indeed,the relationships between IgM and PBC have been a major focus of research on its pathphysiology,which lead to an increased understanding of the abnormal immune responses involved in PBC.This review summarizes data on the biological and clinical significance of Hyper-IgM in PBC,and highlights the potential role of IgM for the diagnosis and treatment of PBC.
2.Yeast expression and application of an antifreeze protein from the desert beetle Microdera punctipennis.
Shanshan MENG ; Wenping CAI ; Ji MA
Chinese Journal of Biotechnology 2015;31(8):1255-1265
Insect antifreeze protein (AFP) has high antifreeze activity. Antifreeze proteins can be used in cryopreservation of biological tissues and cells. We expressed an antifreeze protein from the desert beetle Microdera punctipennis in yeast and determined the function of the protein at low temperatures. Yeast expression vector, pPIC9K-Mpafp698, was constructed and transformed into Pichia pastoris GS115. The expression of MpAFP698 was induced by methanol, and identified by tricine SDS-PAGE and Western blotting. Mpafp698 gene was inserted into the genome of the host yeast strain GS115, and correctly expressed. Hardly any yeast's own protein was secreted into the media. Cryoprotective experiments showed that MpAFP698 can significantly protect mouse liver as well as other mouse organs from cold damage compared with those in the control of Bovine serum albumin (BSA) addition. Besides, the hemolysis of blood cells protected by MpAFP698 at 4 degrees C was reduced and the survival rate of SF9 cells protected by MpAFP698 after freezing and thawing was increased compared to those of the control with BSA addition. Our results showed that MpAFP698 can be expressed in yeast, which allows a convenient purification of the MpAFP protein that has the cryoprotective effect.
Animals
;
Antifreeze Proteins
;
biosynthesis
;
Blotting, Western
;
Cold Temperature
;
Coleoptera
;
Cryoprotective Agents
;
chemistry
;
Electrophoresis, Polyacrylamide Gel
;
Freezing
;
Insect Proteins
;
biosynthesis
;
Mice
;
Pichia
;
metabolism
;
Sf9 Cells
3.Adult head and face models:localization observation of the angular artery and its clinical significance
Chunxiao MA ; Yuanyuan LIU ; Shanshan REN ; Fang WANG ; Xiaosheng LU
Chinese Journal of Tissue Engineering Research 2015;(18):2906-2910
BACKGROUND:Nasolabial fold flap has been widely used in clinical surgery. The facial artery anatomy has been widely used in clinical research. Angular artery dissection is becoming more and more important to nasolabial groove area surgery, but at present, there is a lack of anatomical analysis of internal angular artery.
OBJECTIVE:To study the anatomy of the angular artery, and to provide anatomical data for protecting the nasolabial flap during surgery.
METHODS:Twenty sides of adult cadaver specimens on head and face were dissected. A reference coordinate system was made based on the line between the connection of two medial angles of eyes (axis X) and the facial midline line (axis Y). The location of the angular artery was measured taking A-F as reference points.
RESULTS AND CONCLUSION:(1) The slant angles of the angular artery on BC section, CD section, DE section and EF section were (11.1±4.3)°, (34.1±8.8)°, (21.5±10.5)°, and (17.0±4.7)°, respectively. (2) The angular artery sourced from facial artery was more than it sourced from ophthalmic artery. The diameter of right blood vessel was larger than that of left side. (3) The angular artery sourced from ophthalmic artery comes from the location which extended 8.1 mm to both sides from the point which was 10 mm up from the intersection of facial medial angle of eyes connection and midline. The blood vessel diameter of the starting point was (0.7±0.2) mm. The whole range was 20.1 mm. (4) The angular artery sourced from facial artery comes from the location which extended 25.8 mm to both sides from the point which was 40 mm down to the intersection of facial medial angle of eyes connection and midline. The blood vessel diameter of the starting point was (0.9±0.3) mm. Point to the wing of nose the lateral distance was (5.0±1.2) mm. The whole range was 68.7 mm. The surface projecting of angular artery coming from research results provided anatomic basis for surgery of nasolabial flap.
4.Efficacy of different duration of low-carbohydrate diet intervention on glycosylated hemoglobin in ;type 2 diabetes patients:a Meta analysis
Yanyun ZHANG ; Shanshan MA ; Lili WANG ; Chenglin ZHANG ; Xiaohua WANG
Chinese Journal of Practical Nursing 2016;32(4):317-320
Objective To evaluate the efficacy of different duration of low-carbohydrate diet (LCD) intervention on glycosylated hemoglobin (HbA1c) in type 2 diabetes mellitus (T2DM) patients. Methods Randomized controlled trials (RCTs) of LCD intervention in T2DM patients were collected in the databases such as MEDLINE, PubMed, OVID, China National Knowledge Infrastructure, China Scientific Journal Database by VIP, Wanfang database, et al.Data were analyzed by RevMan 5.3 version. Results Eight RCTs were included. The results of Meta-analysis indicated that the effects of lowering HbA1c by LCD intervention for three (Z=2.28, P<0.05) and six months (Z=14.99, P<0.01) were better than other diabetes diets, but there was no significant statistical difference between one (Z=0.65, P=0.51) and two years (Z=1.62, P=0.10). Conclusions Hypoglycemic effect of short-term LCD was better than other diabetes diets, but long-term effect was similar between them. LCD was a therapeutic diet suitable for T2DM patients.
5.Analysis of 114 Cases of Serious ADR in Our Hospital
Hongyan ZHUANG ; Shanshan LIU ; Hongbing HAO ; Wei GUO ; Xin MA
China Pharmacy 2016;27(35):4933-4936
OBJECTIVE:To investigate the characteristics of serious ADR in psychiatric hospital,and to provide reference for effective and safe use of drugs in the clinic. METHODS:114 cases of serious ADR in our hospital from 2004 to 2015 were ana-lyzed retrospectively in terms of general information,primary disease,route of administration,dosage form,drug types,organs or systems involved in ADR and clinical manifestations,top 5 ADR-inducing drugs and main clinical manifestations,etc. RESULTS:Of 114 serious ADR cases,the number of female was more than that of male(71/43). Patients aged 21-30 years old took up the greatest proprotion(35 cases,30.70%). The main involved primary disease was schizophrenia(54 cases,47.37%). Oral adminis-tration-induced ADR was most common(103 cases,90.35%). The main involved drugs were antipsychotics,occupying the great-est proportion(75 cases,65.79%). Digestive system involved was the most common(53 cases,42.74%), Olanzapine tablets had the highest incidence of serious ADR (23 cases,18.55%),the main clinical manifestation were both abnormal liver function. There were 8 cases of new serious ADR (7.02%). Most of the ADR can be cured after drug withdrawal or countermeasures,no death case was found. CONCLUSIONS:Psychiatric drugs can induce serious ADR as treat psychiatric disease,among which the in-cidence of ADR induced by Olanzapine tablets is the highest and mainly manifestes digestive system symptoms. It is suggested to strengthen serious ADR monitoring to promote the safe use of drugs.
6.Expression of the basic fibroblast growth factor gene after being transfected into rat bone marrow mesen-chymal stem cells
Yue ZHAO ; Lin MA ; Shanshan PENG ; Xin GAN
Journal of Medical Postgraduates 2015;(12):1246-1251
Objective Bone marrow mesenchymal stem cells ( BMSCs) , a kind of stem cells with multiple differentiation po-tentials, exist in the bone marrow and other organizations.This study aimed to investigate the repairing effect of the exogenous basic fi-broblast growth factor ( bFGF) against chronic obstructive pulmonary disease ( COPD) and its action mechanism, and to determine the expression of the bFGF gene in transfected rat BMSCs. Methods BMSCs were isolated, cultured and identified.The recombinant plasmid bFGF-pcDNA3.1 was constructed and sequenced.Liposome-mediated bFGF-pcDNA3.1 plasmid was transfected into the BM-SCs of the rat (bFGF-pcDNA3.1 transfection group), liposome-mediated pcDNA3.1 transfected into the BMSCs (pcDNA3.1 transfec-tion group) , and untransfected BMSCs used as the control.G418 screening was performed for 14 days.The gene and protein expres-sions of bFGF were determined by qRT-PCR and Western blot. Results The full-length sequence of the bFGF gene was consistent with that of the GenBank.The expression of the bFGF gene was significantly higher in the bFGF-pcDNA3.1 transfection group (7.028 ±0.568) than in the pcDNA3.1 transfection group (1.000 ±0.082) and the non-transfection control (1) (P<0.01), but with no statistically significant difference between the latter two groups (P>0.05).The expression of the bFGF protein was also re-markably higher in the bFGF-pcDNA3.1 transfection group (1.017 ±0.054) than in the pcDNA3.1 transfection group (0.217 ± 0.009) and the non-transfection control (0.165 ±0.013) (P<0.05), with no statistically significant difference between the latter two groups (P>0.05). Conclusion Mediated by the liposome reagent, the recombinant eukaryotic expression vector bFGF-pcD-NA3.1 can be transfected into rat BMSCs and expresses the bFGF gene and protein.
7.The influence of bFGF gene transfected BMSCs on inflammatory cytokines expression of COPD rat
Peng WANG ; Qiang NIE ; Lin MA ; Shanshan PENG ; Xin GAN
The Journal of Practical Medicine 2016;32(3):355-358
Objective To study the influence of bFGF gene transfected bone marrow-derived mesenchymal stem cells (BMSCs) on the inflammatory cytokines of COPD rat. Methods The BMSCs were separated from SD rat and cultured and then bFGF gene was imported to BMSCs by liposome transfection method. The samples were prepared into six groups: normal control group, COPD group (A), BMSCs group (B), pcDNA3.1-BMSCs group (C), bFGF-pcDNA3.1-BMSCs group (D), and bFGF group (E). The expressions of TNF-α and IL-1β by QRT-PCR were detected. Results Compared with COPD group, TNF-α and IL-1β genes from groups B to D dropped significantly (P < 0.05). The changes of TNF-α and IL-1β genes among groups B to D showed no significant difference (P > 0.05). Conclusion BFGF transfected BMSCs, sample BMSCs and pcDNA3.1 transfected BMSCs can inhibit the expression of inflammatory cytokines of TNF-α and IL-1β, but there is no obvious advantage in comparison to bFGF transfected BMSCs and sample BMSCs in respect of inhibiting the expression of inflammatory cytokines of TNF-α and IL-1β.
8.Runx2 over-expression effect on expression of osteogenesis related genes in osteogenic differentiation of human umbilical cord blood mesenchymal stem cells
Dakai ZHOU ; Huining LI ; Shanshan MA ; Tian CHENG
Chinese Journal of Tissue Engineering Research 2017;21(9):1444-1449
BACKGROUND: Runx2 plays a central role in osteogenic differentiation, which is of important significance in new bone formation.OBJECTIVE: To observe the effect of Runx2 over-expression on osteogenic differentiation of human umbilical cordblood mesenchymal stem cells.METHODS: Runx2 was generated by RT-PCR and the recombinant adenovirus (pAd-Runx2) was constructed. The viraltiter was determined by air dilution method. After being transfected into human umbilical cord blood mesenchymal stemcells, the green fluorescence was observed under fluorescence microscope. Real-time fluorescent quantitative PCR andwestern blot were used to detect mRNA expression changes of osteogenesis related genes, Runx2, OCN, BMP-2, ALP,in human umbilical cord blood mesenchymal stem cells at 1, 3, 7 and 14 days after transfection.RESULTS AND CONCLUSION: The recombinant adenovirus was successfully constructed and the titer was1.7×1010 pfu/L. After infection by pAd-Runx2, human umbilical cord blood mesenchymal stem cells expressed greenfluorescence protein clearly under the fluorescence microscope. Cells in the transfected group differentiated intoosteoblast-like cells, and those in the control group stayed the same as pre-infected. The expression of Runx2, OCN,BMP-2 and ALP in the transfected group increased over time to some extent, but these changes were not detected in thecontrol group. These findings indicate that Runx2 over-expression can promote the osteogenic differentiation of humanumbilical cord blood mesenchymal stem cells.
9.Determination of Mycoplasma genitulium infection in patients with chronic non-bacterial prostatitis and its significance
Shuxia ZHONG ; Shanshan LI ; Chaoying ZHANG ; Cuiyang ZHAO ; Lei MA
Journal of Jilin University(Medicine Edition) 2006;0(01):-
Objective To study the action of Mycoplasma genitulium (Mg) in pathogenesis of chronic non-bacterial prostatitis. Methods Mg culture and polymerase chain reaction (PCR) were perfomed on prostatic secretion specimens from 987 patients with chronic non-bacterial pristatitis and 125 healthy men. Results ①The positive rate of Mg in the prostatic secretion from the 987 patients was 31.5% (302/987). Among the 302 positive cases, 153 cases had other pathogens, of which Ureaplasma urealyticum (Uu) was the most (128). ②In the controls, the positive rate of Mg was 2. 53% (3/125). The difference between the two groups was statistically significant (X2=44. 32,P
10.Role of spinal sigma-1 receptors in maintenance of bone cancer pain in rats: the relationship with ERK
Chenchen WANG ; Shanshan ZHU ; Yuan HAN ; Xiao LI ; Wenwen MA
Chinese Journal of Anesthesiology 2014;34(7):796-799
Objective To evaluate the role of spinal sigma-1 receptors in the maintenance of bone cancer pain (BCP) in rats and the relationship with extracellular signal-regulated kinase (ERK).Methods Part Ⅰ Twenty-four female Sprague-Dawley rats,weighing 180-220 g,were randomized into 2 groups using a random number table:sham operation group (S group,n =4) and BCP group (n =20).BCP was induced by inoculating Walker 256 mammary gland carcinoma cells into the medullary cavity of the right tibia.Four rats were sacrificed on day 10 after inoculation in S group or on day 3,5,7,10 and 14 after inoculation in BCP group,and the L4-6 segments of the spinal cord were removed to measure the expression of sigma-1 receptors by Western blot.Part Ⅱ Forty female Sprague-Dawley rats,weighing 180-220 g,were randomized into 4 groups (n =10 each) using a random number table:sham operation group (S group),sigma-1 receptor inhibitor BD1047 group (BD group),BCP group,and BCP + BD1047 group (BCP + BD group).On day 10 to 14 after inoculation,normal saline 20 μl was injected intrathecally once a day in S and BCP groups,or BD1047 120 nmol/20μl was injected intrathecally once a day in BD and BCP + BD groups.Mechanical paw withdrawal threshold (MWT) to yon Frey filament stimulation was measured one day before inoculation,on day 3,5 and 7 after inoculation,and on day 10,12 and 14 after administration.After measurement of MWT on day 14 after inoculation,the rats were sacrificed and the L4-6 segments of the spinal cord were removed to determine the expression of phosphorylated ERK (p-ERK) by Western blot.Results Part Ⅰ Compared with group S,the expression of sigma-1 was significantly up-regulated and peaked on day 10 after operation in group BCP.Part Ⅱ Compared with S group,no significant changes were found in MWT and p-ERK expression at each time point in BD group,and MWT was decreased and p-ERK expression was up-regulated in BCP and BCP + BD groups.Compared with group BCP,after intrathecal injection of BD1047,MWT was significantly increased and the expression of p-ERK was down-regulated in BCP + BD group.Conclusion Spinal sigma-1 receptors are involved in the maintenance of BCP in rats possibly through promoting phosphorylation of ERK.