BACKGROUND:Pain after hip replacement is a difficulty for orthopedic surgeons,and revision surgery might result in severe wound for patients undergoing hip replacement.Extracorporeal shock wave(ESWT) is certificated effective for relieving pain.OBJECTIVE:To evaluate the effects of ESWT on the pain after hip replacement.DESIGN,TIME AND SETTING:Case self-control,double-blind evaluation.The experiment was performed at the Department of Orthopaedics,Zhongnan Hospital,Wuhan University between June 2004 and February 2008.PARTICIPANTS:Fifty-one cases(61 hips) undergoing total hip replacement with pain were selected,including prosthetic loosening(cemented:16 cases,20 hips;non-cement:12 cases,16 hips),ectopic ossification(8 cases,10 hips),acute infection(4 cases,4 hips),chronic infection(5 cases,5 hips),soft tissue aseprtic inflammation(4 cases,4 hips) and acetabulum dislocation(2 cases,2 hips).METHODS:These patients received sessions of ESWT(4 000 shocks,3 times/s),with an energy ranging from 0.54 to 1.06 MJ/mm2,once a week for 8-12 weeks.MAIN OUTCOME MEASURES:Visual analog scale(VAS),Harris hip scores,diversity of the sclerotin surrounding the prosthesis(X-ray).RESULTS:The 51 patients were followed up for 18-64 months(4.1years in average),and 2 died during this period.The scores of VAS in non-cement,chronic infection,ectopic ossification and soft tissue aseprtic inflammation groups were significantly decreased than before treatment(P

Objective To cxplore the variation of the range of motion (ROM) of operative level after different heights of artificial cervical disc replacement,and to provide guidance for clinical work in selecting appropriate height of artificial cervical disc prosthesis.Methods The preoperative cervical anteroposterior and lateral Ⅹ-rays of 9 fresh male cadaveric cervical spine specimens were obtained to measure the intervertebral height of C5-6,and 3 screened specimens with the height of about 5 mm were included in the experiment.The experiment was designed to test self-control,and other four groups of cervical specimens including intact group,appropriate height (5 mm) of C5.6 artificial cervical disc replacement group,1mm increased (6 mm) group and 2 mm increased (7 mm) group were made biomechanical test sequentially.The specimens were fixed to the cervical three-dimensional movement machine,with a 75 N follower load and pure moments of 2 Nm for flexion/extension 、left/right bending and left/right axial rotation,to measure the ROM of operative level under the condition of changes in 0.2 Nm/s.Results There were no significant differences in the ROM of flexion/extension,lateral bending and axial rotation between 5 mm group and intact group;the ROM of flexion/extension、lateral bending and axial rotation in 6 mm group increased compared with 5 mm group,but the difference was not statistically significant;the ROM of flexion/extension in 7 mm group was significantly less than that of intact,5mm and 6 mm group (9.5°± 1.0° vs 12.5°±0.9°、11.3°±0.8°、11.6°±0.9°),but significantly greater in axial rotation than 6 mm group (10.4°±1.4°vs 8.6°±0.3°),and there was no significant difference in lateral bending compared with other 3 groups.Conclusion 2 adjacent heights of cervical disc prostheses are implantedsuitably when testing the mold of disc prosthesis,the choice of cervical disc prosthesis with 1 mm increased can improve the ROM of operative level to some extent;while the height with 2 mm increased can lead to the ROM of flexion/extension at the operative level reduced,but the ROM of rotation shows an increasing trend.

Objective To express the receptor binding domain (RBD) protein of the Middle East respiratory syndrome coronavirus (MERS-CoV) and to characterize the antigenicity of the purified recombi-nant protein. Methods The codon-optimized gene encoding the RBD protein of MERS-CoV was synthesized and then cloned into the pET30a ( +) vector to construct the recombinant expression plasmid. The trans-formed E. coli BL21 (DE3) strains carrying expression plasmid were induced by IPTG under different condi-tions. The expressed products were purified by using nickel affinity chromatography and further analyzed by SDS-PAGE and Western blot assay. Indirect ELISA was performed to analyze the antigenicity and specificity of RBD proteins expressed in prokaryotic expression systems in human serological test. Results The recom-binant RBD proteins were mainly expressed as conclusion body in an optimal induction condition of 37℃ and 0. 5 mmol/ L IPTG for 4 h. The high purified recombinant RBD proteins were obtained through denaturation and renaturation with a relative molecular mass of about 29×103 . Results of the Western blot assay showed that the recombinant RBD proteins could have specific reaction with the serum samples collected form mice with MERS-CoV infection. Indirect ELISA revealed that the RBD proteins expressed in the prokaryotic ex-pression system showed better sensitivity and specificity in the detection of antibodies against MERS-CoV in human serum samples. Conclusion This study reported the prokaryotic expression and purification of RBD protein of MERS-CoV for the first time, which might pave the way for further investigation on immunological detection of MERS-CoV and development of vaccines against MERS-CoV infection.

Five groups were assigned to include intrahepatic cholangiocarcinoma ( ICC, n=30 ) , liver cirrhosis (LC,n=30),metastatic carcinoma (MCA,n=30) and 30 healthy subjects.The serum level of GPC3 was measured by a sandwich method of enzyme-linked immunosorbent assay ( ELISA ) and alpha-fetoprotein (AFP) by microparticle enzyme immunoassay (MEIA).The serum levels of GPC3 and AFP were significantly higher than those of other groups (P<0.05).At a cut-off value of 3.5μg/L,the sensitivity and specificity of GPC3 in the diagnosis of HCC was 83.3%and 76.7%respectively.The sensitivity of combined measurement of GPC3 and AFP was better than GPC3 or AFP alone.Detectable GPC3 was significantly correlated with the presence of viral hepatitis markers and tumor size.However there was no obvious difference in tumor thrombi in portal vein ( PVTT), tumor number, age, gender or hepatic function of HCC.Thus,as a sensitive serum diagnostic marker for HCC ,GPC3 may be a good supplement to AFP in differentiating HCC from non-malignant chronic liver diseases and other liver cancers.

[Objective] To investigate the effects and mechanisms of Nrf2-ARE (nuclear factor erythroid-2 related factor-anti-oxidant response element) pathway on uremic serum-mediated endothelial dysfunction in human aortic endothelial ceils.[Methods] Human aortic endothelial cells were incubated in endothelial cell medium containing 10％ normal serum,10％ non-diabetic nuremic serum or 10％ diabetic uremic serum respectively,and 20 μmol/L tertiary butyl hydroquinone (tBHQ) were pretreated with cells to active Nrf2-ARE pathway.The cells apoptosis rate were measured by flow cytometry,and the synthesis of NO was detected by flow cytometry and immune fluorescent confocal,while the expression of P-eNOSer1177/eNOS,and quinone oxidoreductase-1 (NQO1) were measured by western blotting.The levels of malondialdehyde,superoxide dismutase,eatalase,and glutathione in these cells were also measured with kits.[Results] Aortic endothelial cells incubated with uremic serum had a higher level of apoptosis rate and MDA (P ＜ 0.05),and a lower level of NO systhesis,P-eNOSSer1177/eNOS expression,CAT,SOD,GSH (P ＜ 0.05).Pretreated with tBHQ can reduce the apoptosis rate and MDA level (P ＜ 0.05),improve the amount of NO systhesis,the expression of P-eNOSSer1177/eNOS,the levels of CAT,SOD,and GSH in these cells (P ＜ 0.05).[Conclusion] Activation of Nrf2-ARE pathway can improve endothelial dysfunction in aortic endothelial cells induced by uremic serum,and its mechanism might be related with enhancement of the antioxidant stress.

Objective To evaluate the prevalence and clinical features of human Coronavirus OC43 (HCoV-OC43) in adult patients with acute respiratory infections (ARI) in Beijing.Methods Nasopharyngeal swab specimens were collected from 559 adult patients with ARI.HCoV-OC43 infection was detected using two sets of OneStep reverse transcription polymerase chain reaction (OneStep RT-PCR),which targeted the spike and nucleocapsid coding region.Results The prevalence of HCoV-OC43 was 12.52％ (95％ CI:9.78％-15.26％),and the epidemic peak was in autumn.The dominant clinical presentations of HCoV-OC43 were fever,sore throat,headache,cough,nasal stuffiness,nasal discharge,and so on; 8 patients showed gastrointestinal symptoms such as diarrhea and vomiting.Statistically,nasal stuffiness was the most representative clinical presentation.Coinfection of HCoV-OC43 with other respiratory viruses was shown to be 35.71％ (25/70,95％ CI:24.49％-46.93％).Conclusion With sensitive molecular detection techniques and nasopharyngeal swabs,high rate of HCoV-OC43 infection was achieved in adult patients with ARI in Beijing.

OBJECTIVETo develop an one-tube multiplex RT-PCR assay for simultaneous detection of six human coronaviruses (HCoVs).

METHODSGenbank sequences of six human coronaviruses, including HCoV-NL63, HCoV-229E, SARS-CoV, HCoV-OC43, MERS-CoV, and HCoV-HKU1, were included as reference sequences. Primers were designed based on multiple alignment of reference sequences, targeting the conserved regions of each species of HcoV. Virus strains and nucleic acid preserved in our lab were used as template in developing this automatic-electrophoresis-based one-tube multiplex RT-PCR assay. Detection limits and reproducibility were also evaluated with these templates. Samples with infection of other respiratory viruses preserved in our lab were used to evaluate specificity of this assay. Finally, we tested this assay with 140 clinical samples that were validated by real-time PCR in parallel.

RESULTSThis automatic-electrophoresis-based multiplex RT-PCR assay was able to detect six human coronaviruses simultaneously. All positive samples in this study were detected with at least one specific fragment of anticipated length (195, 304, 332, 378, 415, 442 bp) . No fragment was detected in negative controls. Detection limits of 1.0×10(1-1.0)×10(2) copies/µl were achieved in tests of single virus. No cross reaction was observed with other respiratory viruses. This multiplex RT-PCR assay showed the same sensitivity and specificity to that of individual real-time RT-PCR validated with clinical samples. Both methods detected 28 positive samples (20%) .

CONCLUSIONSSix HCoVs can be detected in one tube by this novel multiplex RT-PCR assay with high sensitivity and specificity.

Objective To investigate the genetic diversity differences of human parvoviruse B19 and parvovirus 4 (PRVA4) in Tibet and Han population in China.Methods Phylogenetic analysis was performed on genome fragments of B19 or PARV4 obtained from the blood samples of Tibet and Han population in China by using a PCR followed by sequencing.Results Ten partial VP1 fragments of B19 (2 from Tibet,3 from Sichuan,5 from Zhejiang) and 10 partial ORF1 fragments of PAV4 (2 from Tibet,2 from Sichuan,1 Yunnan,5 from Zhejiang) were obtained.Phylogenetic analysis indicated that different B19 subgroup circulates in Tibet and Han population although they belong to the same 1A subtype.While the gene evolution of PAV4 is very conserved between the Tibet and Han population in China.Conclusion These studies on genetic diversity of B19 in different Chinese population provide a way for detection and prevention of B19 human parvovirus infection.

ObjectiveTo analyze the characteristics of death and premature death of 4 major chronic diseases （cardiovascular and cerebrovascular diseases， malignant tumors， chronic respiratory diseases and diabetes） in Taizhou City from 2011 to 2018，and provide data basis for the government to formulate chronic disease prevention planning. MethodsThe death data of household registration residents in Taizhou City from 2011 to 2018 were derived from the Chronic Disease Surveillance Information Management System in Zhejiang Province. The death toll ratio of chronic diseases， the mortality rate of chronic diseases， the probability of premature death of chronic diseases were analyzed. The standardization rate was calculated six times in 2010. Population composition of the census. The Joinpoint Regression Program 4.2 software was used for calculating annual percent change （APC） and its statistical test results. ResultsFrom 2011 to 2018， there were 231 724 chronic disease deaths in Taizhou City， with a mortality rate of 486.52/10⁵ and a standardized mortality rate of 381.55/10⁵. The proportion of chronic disease deaths to total deaths was 79.89%， of which males were higher than females and rural areas were higher than urban areas.From 2011 to 2018， the standardized mortality and early death probability of cardiovascular and cerebrovascular diseases， malignant tumors and chronic respiratory diseases in Taizhou showed a downward trend （P<0.05）， the standardized mortality of diabetes （P=0.46） and the early death probability （P=0.22） did not decline， and the mortality of all age groups of the above four types of chronic diseases in rural areas was higher than that in urban areas. The mortality of the four types of chronic diseases from high to low are cardiovascular and cerebrovascular diseases， malignant tumors， chronic respiratory diseases and diabetes， and the mortality tends to increase with age. From 2011 to 2018， the probability of premature death from four types of chronic diseases in Taizhou City showed a downward trend， from 13.49% in 2011 to 10.49% in 2018， with an average annual decrease of 2.97%. The difference was statistically significant （t=‒5.83，P<0.05）. ConclusionChronic disease death is the main cause of death in Taizhou City. In order to reduce the mortality rate of chronic diseases， effective prevention and control measures for chronic diseases should be carried out， especially the prevention and control of diabetes and male chronic diseases.