2.Primary culture of human vascular smooth muscle cells and their calcification mode
Xiao-zhong, CHENG ; Ting, SONG ; Bei, HUANG ; Hui, ZHONG
Bulletin of The Academy of Military Medical Sciences 2010;34(1):37-39
Objective To establish a calcification mode in vitro of human vascular smooth muscle cells (HVSMCs) induced by β-GP. Methods Primary HVSMCs were obtained from human embryo by plant method and confirmed by stain with α-sm-actin antibody. The cells after 4-6 passages were divided into two groups.The control group was incubated with normal DMEM medium while the calcification group was incubated with the medium containing 10 mmol/L β-glycerophosphate for 10 days.Calcification was confirmed by alizarin red S stain and alkaline phosphatase(ALP) assays. Results The primary cells observed by S-P stain were positive and the cells after being stained were pale yellow. After being induced with β-GP, the cells of calcification group began concentric growth and formed vesicles. Alizarin red S staining showed that the reaction of calcifying nodules was red,ALP activity was higher than that of controls at various time points(4 d,6 d,8 d and 10 d,P<0.01 ).Conclusion The HVSMCs could be induced into calcification in vitro by β-GP, and this model contribates to further studies of vascular diseases.
3.Cemented artificial femoral head arthroplasty treats elderly unstable intertrochanteric fracture
Shigao CHENG ; Wanchun WANG ; Yang XIAO ; Tunsheng HUANG ; Zhong XIE
Chinese Journal of Tissue Engineering Research 2015;(52):8373-8378
BACKGROUND: Many studies have suggested that artificial joint replacement has a greater advantage in the treatment of cases of elderly osteoporosis, comminuted fractures and unable to internal fixation or failure of internal fixation. OBJECTIVE:To analyze the curative effect of cemented artificial femoral head arthroplasty for treatment of elderly unstable intertrochanteric fracture. METHODS: Totaly 18 patients with unstable intertrochanteric fracture, aged>75 years, were enroled and underwent cemented artificial femoral head arthroplasty. At 3 months after replacement, the curative effect was evaluated according to the Harris hip joint function score method. The early complications after replacement and long-term complications of prosthesis were observed by folow-up. RESULTS AND CONCLUSION:Al these 18 patients were folowed up for 13 to 34 months, and had no infections, pressure sores, femoral shaft fractures and other complications. X-ray films showed that the frauture healed wel, without prosthesis infection, dislocation, loosening, sinking and breaking. After 3 months of replacement, al patients returned to pre-injury level of walking function. The curative effect was excelent for eight cases, good for seven cases, fair for two cases, poor for one case. These results demonstrate that cemented artificial femoral head arthroplasty for treatment of elderly unstable intertrochanteric fracture has good biocompatibility and stability, and can restore the limb function of patients. 
4.Comparison of mild hypothermic and moderate hypothermic cardiopulmonary bypass in pediatric cardiac surgery
Wei CHENG ; Yingbin XIAO ; Lin CHEN ; Qianjin ZHONG ; Xuefeng WANG ; Mei LIU ; Li PENG ; Wei HU
Journal of Third Military Medical University 2001;23(5):508-510
Objective To compare the effects of mild hypothe rmic cardiopulmonary bypass (CPB) and moderate hypothermic cardiopulmonary bypas s in pediatric cardiac surgery. Methods A total of 118 cas es of less than 3 years of age that had undergone open heart surgery were review ed, in which 46 patients received moderate hypothermic CPB(group 1) and 72 patie nts received mild hypothermic CPB(group 2). The CPB duration, incidence of low c ardiac output and postoperative concentration of CK-MK, etc, were compared with each other betwee n the two groups. Results Duration of bypass and postoperative mechanical respiratory assistance of group 2 was shorter than that of group 1 ( P<0.05). Transfusion requirements, incidence of low cardiac output syndrome, concentration of CK-ME and percentage of metabolic acidosis were lower in grou p 2 than in group 1 (P<0.05), while the index of oxygenation was higher in g roup 2(P<0.05). Conclusion The mild hypothermic CPB is saf er and more effective and therefore is superior to moderate hypothermic CPB in p ediatric cardiac surgery.
5.Effect of pulsed electromagnetic fields of different daily treatment durations on biomechanical properties of femur in ovariectomized rats
Cheng-Qi HE ; Wei WANG ; Deng XIAO ; Lin YANG ; Zhong-Jie LEI ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(08):-
Objective To observe the effect of pulsed electromagnetic fields(PEMFs)of different daily treatment durations on biomechanical properties of femur in ovariectomized rats,so as to find out the optimal daily treatment time.Methods Fifty female Sprague-Dawley rats were randomly divided into five groups:(1)SHAM control(no PEMFs treatment),(2)OVX control(no PEMFs treatment),(3)OVX I(PEMFs treatment at 8Hz fre- quency with 3.8 mT intensity,20 min daily for 30 days),(4)OVX 11(PEMFs treatment at 8 Hz frequency with 3.8 mT intensity,40 min daily for 30 days),and(5)OVXⅢ(PEMFs treatment at 8Hz frequency with 3.8 mT in- tensity,60 rain daily for 30 days).All the rats were subject to bilateral overiectomy except those in the SHAM control group.The biomechanical properties of the femur were assessed after 30 days of PEMFs treatment.Results The values of the parameters of the biomechanical properties obtained with the OVX control group were significantly lower than those of the other 4 groups(P0.05).Conclusion PEMFs of the three different daily treatment durations can maintain the biomechanical properties of the femur in ovariectomized rats.Under certain in- tensity(3.8 mT)and frequency(8 Hz),PEMFs of the three different treatment durations can significantly maintain biomechanical properties of femur in ovariectomized rats approximately to the nomal level,but among the three groups,the difference is not significant.It was shown that exposure to PEMFs for 20 to 60 minutes daily had similar effect of maintaining biomechanical properties of the femur in ovariectomized rats.
6.Neuroprotective effects of erythropoietin on cultured retinal neurocytes by glutamate
hui-ping, YAO ; yi-sheng, ZHONG ; yu, CHENG ; xiao-hong, LIU
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(12):-
Objective To investigate whether erythropoietin(Epo) is potentially beneficial in protecting cultured retinal neurocytes.Methods Primary isolated retinal nerve cells were cultured.Expressions of Epo and EpoR protein in cultured retinal neurocytes were decected by immunohistochemical analysis.Survival of cultured neurocytes that were incubated in the presence of Epo or glutamate in the presence or absence of Epo were estimated by determining the activity of their mitochondrial dehydrogenases using the MTT assay.Results Epo and EpoR protein were expressed on the cultured retinal neurocytes.The presence of different concentrations of Epo did not improve the survival of retinal neurocytes,and Epo could prevent glutamateinduced toxicity. Conclusion Epo is beneficial in protecting mixed cultured retinal neurocytes from glutamate-induced cytotoxicity.
7.Analysis of iodized salt monitoring results in Chongqing Municipality from 2001 to 2009
Bang-zhong, XIAO ; Jing, CHEN ; Cheng-guo, WU ; Ya-lin, CHEN ; Xing-jian, LUO
Chinese Journal of Endemiology 2011;30(4):416-419
Objective To monitor the quality changes of iodized salt and analyze its impact factor in Chongqing between 2001 and 2009. Methods Salt samples were collected according to the east, west, south,north and center locations in iodized salt production, wholesale and household sectors. Two units in iodized salt production and wholesale segment were sampled from north, south, east and west places and only 1 unit was sampled from the central place. Nine samples were collected every month in each place. If the place had less than 9 units, and then taken all the units. About resident household, 2 townships were sampled from north, south, east and west places, and 1 township was sampled from the central place, then 20 samples were collected from each township. Iodine content was detected by oxidation-reduction assay. The index of mean iodine, qualified rate from factories and wholesale, coverage rate and taking rate of qualified iodized salt in residents were calculated.Significance was analyzed by trend test, analysis of variance and X2 test. Results The qualified rate of iodized salt from the manufacturers was 92.9%(13/14) in 2001 and the rate was 100.0% each year from 2002 to 2009. The qualified rates of iodized salt from the wholesale were 88.7%(282/318) - 99.8%(431/432). The rates of 2001 and 2002 were lower than that of other years(X2 = 4.98 - 45.69, all P< 0.05 or < 0.01). The coverage rate and taking rate of qualified iodized salt in residents were 94.2% (11 154/11 841 ) - 98.9% ( 14 061/14 217), 83.5% (9 887/11 841 ) -95.8% (13 449/14 039), respectively. The rates showed an increasing tendency (F = 9.27, 26.39, all P < 0.05).The districts(counties) with qualified iodized salt consumption rate > 90% kept increasing. The mean iodine from the manufacturers and wholesale were 29.71 - 36.25, and 31.26 - 36.13 mg/kg, respectively. The iodine level showed a descending trend(F = 35.45, 140.59, all P < 0.01 ). The mean iodine level from the inhabitants were 28.84 - 30.98 mg/kg which remained stable (F = 3.05, P > 0.05 ). The iodine level from manufacturers, wholesale to inhabitants showed an descending trend(F = 38.46 - 671.23, all P < 0.01 ). Conclusions The surveillance results of iodized salt shows an increasing tendency in quality of iodized salt, eoverage rate and taking rate of qualified iodized salt. Factors that affect the quality of iodized salt is that the enterprise does not add iodine to salt strictly by the standard.
8.Construction and expression of the recombinant plasmid pET28α-Sj26GST-Sj32 of Schistosoma japonicum in Escherichia coli BL21(DE3)
Wen-gui, LI ; Bang-zhong, XIAO ; Xing-jian, LUO ; Ya-tang, CHEN ; Cheng-guo, WU
Chinese Journal of Endemiology 2011;30(2):152-157
Objective To construct and express the recombinant plasmid pET28α-Sj26GST-Sj32 of Schistosoma japonicum(Sj) in Escherichia coli BL21 (DE3). Methods Total RNA was extracted from Sj adult worms by ultrasound-breaking, Sj26GST and Sj32 antigen gene was respectively amplified by RT-PCR from the total RNA; Sj26GST-Sj32 fusion gene obtained with gene splicing by overlap extension(SOEing) was cloned into prokaryotic expression plasmid pET28α and transformed into Escherichia coli BL2 (DE3) to construct pET28α-Sj26GST-Sj32;BL21 (pET28α-Sj26GST-Sj32) was induced with isopropyl-β-D-thiogalactopyranosid (IPTG), and the expressed products were analyzed and identified by sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE)and Western blotting. Results The 1991 bp Sj26GST-Sj32 fusion gene was successfully amplified by gene SOEing and cloned into pET28α by restriction analysis and PCR identification, the recombinant plasmid pET28α-Sj26GST-Sj32 was successfully constructed; the relative molecular mass of the expressed recombinant protein was approximately 69 × 103 by SDS-PAGE, and the amount of the expressed protein was 25% of the total bacterial proteins; the fusion protein could be recognized by sera from rabbits infected with Sj by Western blotting.Conclusions The recombinant plasmid pET28α-Sj26GST-Sj32 is successfully constructed and highly expressed in Escherichia coli in fused form with His-tag, and the expressed fusion protein shows specific antigenicity.
9.Construction and expression of the recombinant plasmid pET32α-Sj26GST of Schistosoma japonicum in Escherichia coli BL21(DE3)
Wen-gui, LI ; Bang-zhong, XIAO ; Xing-jian, LUO ; Ya-tang, CHEN ; Cheng-guo, WU
Chinese Journal of Endemiology 2010;29(3):287-291
Objective To construct and express the recombinant plasmid pET32α-Sj26GST of Schistosoma japonicum(sj)in Escherichia coli(E.coli)B121(DE3).Methods The total RNA was extracted from sj adult worms by ultrasound-breaking,Sj26GST antigen gene was amplified by RT-PCR from the total RNA,then cloned into prokaryotic expression plasmid pET32α(+) and transformed into E.coli B12(DE3)to construct pET32α-Sj26GST;BL21(pET32α-Sj26GST)WaS induced with isopropyl-β-D-thiogalactopyranosid(IPTG),and the expressed products were analyzed and identified by SDS-PAGE and Western blot.Results The 676 bp Sj26GST gene was successfully amplified by RT-PCR and cloned into pET32α(+)by restriction analysis and PCR identification,the recombinant plasmid pET32α-Sj26GST was successfully constructed;the relative molecular mass of the expressed recombinant protein was approximately 49×103 by SDS-PAGE,and the amount of the expressed protein was 24%of the total bacterial proteins;the fusion protein could be recognized by sera from rabbits infected with sj by Western blot.Conclusions The recombinant plasmid pET32α-Sj26GST is successfully constructed and highly expressed in E.coli in fused form with Trx-tag and His-tag,and the expressed fusion protein shows specific antigenicity.
10.Dynamic Distribution of Shrimp White spot syndrome virus (WSSV) Qingdao Strain in Crawfishes by Dot Hybridization
Jian-zhong, ZHU ; Xiao-Qin, XIA ; Cheng-ping, LU ; Fu-sheng, GUO
Virologica Sinica 2001;16(1):92-95
A fragment sized 400bp of White spot syndrome virus(WS SV,formerly de signated NOSV),recovered from recombinant plasmid pAFD, was labeled with Digox igenin as a probe to detect dynamic distribution of WSSV within 120h and 72h in crawfishes(Cambarus proclarkii) inoculated WSSV by oral taking and injecti on r espectively. Stomach epithelium, intestine epithelium, heart, gill, haemolymph, muscle, hepatopancreas, hypoderm, connective tissue and ovary of infected crawfi shes were examined for WSSV. In both groups, WSSV was first detected in heamoly mph at 12h p.i. and then disappeared. Again it was detected at 96h p.i. only in oral infection group and maintained till 120h p.i., but it didn't appear at 72h p.i. in injection group. WSSV in heart, muscle was detected at 36h p.i. in oral infection group and 24h p.i. in injection group respectively, and then increased generally. In addition, WSSV in intestine epithelium, connective tissue, ovary of oral infection group and intestine epithelium, hypoderm, ovary of injection g roup could also be detected. In dead crawfishes after 120h and 72h p.i. in two groups, WSSV could be detected in all the examined tissues and it demonstrated t hat systemic infection occurred in the animales. The tissue containing more amo unts of WSSV was hypoderm in oral infection group, while intestine epithelium, g ill, hypoderm, ovary in injection infection group. It deduced that WSSV first a ppears in haemolymph and then goes into heart, muscle and other tissues and prol iferates in them. Once again, WSSV is released into heamolymph resulting in syst emic infection till crawfishes' death.