OBJECTIVE:To observe clinical efficacy and safety of flupirtine maleate for pain caused by acute lumbar sprain. METHODS:60 patients with acute lumbar sprain were selected and divided into trial group and control group according to even and odd-numbered admission order. Trial group received flupirtine maleate capsule,1 piece/time,3 times/d;control group was giv-en codeine sustained-release tablet,2 tablets/time,2 times/d. The VAS score,clinical efficacy and ADR were compared between 2 groups. RESULTS:The VAS score of treatment group after treatment was significantly lower than that of control group,with statis-tical significance(t=2.375,P=0.013). The clinical efficacy of trial group was significantly higher than that of control group,with statistical significance (u=9.431,P=0.024). The ADR of trial group was mild,and there was no significant difference between two groups(χ2=0.131,P=0.717). CONCLUSIONS:Flupirtine maleate has a good clinical efficacy and safety in the treatment of pain caused by acute lumbar sprain.

Objective To explore the effect of hyperbaric oxygenation (HBO) on generation of free radicals and the permeability of blood-brain barrier(BBB) during cerebral ischemia-reperfusion(CIR). Methods Three hundred and twenty Kunming mice were randomly divided into four groups: a sham operation group, a HBO group, a CIR group, and a HBO + CIR group, with 80 mice in each group. Conscious mice were used to establish the CIR model, and 0.25MPa (ATA) HBO were applied 5 times after operations. The activities of SOD, CAT, GSH-PX and the concentration of MDA, EB in the cerebral hippocampal tissues in each group were measured with colorimetry. The cerebral hippocampal tissues were harvested and processed, then observed and compared with transmission electron microscopic observation. Results Compared with the sham operation group, the activities of antioxidation enzymes (GSH-PX,SOD, CAT) in CIR group decreased significantly (P

Objective To clone and express the partial encoding sequence of Mr 70 000 heat shock protein of Cryptosporidium andersoni (CaHSP70) in Escherichia coli and identify the recombinant protein. Methods Total RNA was extracted from oocysts of C.andersoni isolated from Xuzhou, Jiangsu (XZ-BOV). The CaHSP70 gene was amplified by RT-PCR. The PCR product was cloned and then subcloned into pET28a vector, and the recombinant plasmids were transformed into E.coli BL21(DE3) subsequently. The expressed protein induced by IPTG was purified and identified by SDS-PAGE and Western blotting, and was further analyzed by relevant bioinformatics softwares. The specific IgG antibodies in mice immunized by rCaHSP70 were detected by Western blotting and ELISA respectively. Results The deduced amino acid sequence showed to be identical with that of C. andersoni Mr 70 000 heat shock protein (HSP70). The recombinant protein expressed in the form of inclusion body was about Mr 43 000. It could be recognized by anti-His G labeled HRP antibodies and all the sera from mice infected with C. andersoni and children infected with C. parvum as well as sera from mice immunized with rCaHSP70 respectively. The rCaHSP70 possibly had multiple domains and potential antigenic determinants. Phylogenetic analysis showed that XZ-BOV and C. andersoni were in the same clade. ELISA showed that the level of specific antibodies against rCaHSP70 in immunized BALB/c and C57BL/6 mice was significantly higher than that of mice before immunization. Conclusion The recombinant plasmid pET28a-CaHSP70 has been constructed. The purified rCaHSP70 exhibits high antigenicity and seems a potential candidate antigen for immunodiagnosis of cryptosporidiosis.

Objective To perform the cloning of the gene encoding Schistosoma japonicum Chinese-strain hypoxanthine-guanine phosphoribosyltransferase(HGPRT)and its expression in Escherichia coli.MethodsA couple of primers were designed with the BamHI restriction endonuclease site introduced in forward primer and SalI in reverse primer.Total RNA was isolated from adult worms of S.japonicum Chinese-strain(Anhui-strain,Sjc-A)and the SjcHGPRT gene was amplified by reverse transcriptase-polymerase chain reaction(RT-PCR).The PCR product and the prokaryotic expression vector pET28a were digested by both restriction endonucleases BamHI and SalI.The target DNA fragments were purified and cloned properly into pET28a.After identification by en-donucleases digestion,PCR and sequencing,the recombinant plasmid pET28a-SjcHG PRT was transformed into competent E.coli BL21 and expressed in the presence of IPTG.Results pET28a-SjHGPRT was sequenced and shown to be 99% and 83% identical in deduced amino acid sequence to that of S.japonicum Chinese-strain(Hunan-strain,Sjc-H)and S.mansoni HGPRT,respectively.The results of SDS-PAGE and Western blot revealed that the molecular weight of expressed protein was around 30 kDa and could be recognized by anti-His-G-HPR antibody and sera from mice and human with schistosomasis japonica.Conclusion The recombinant plasmid containing SjcHGPRT cDNA is successfully constructed and its expression protein(reSjcHGPRT)is also successfully purified.

Objective Optimal detection of deep hypothermic circulatory arrest (DHCA)-induced early brain injury is important but the effective technique is still not available in the present.The relationship between diffusion wcighted imaging (DW1) and histopathological changes in DHCA-induced piglet brain injury model were analyzed.Methods Eighteen pigs underwent deep hypothermic circulatory arrest and were divided into three groups:group A (n =6) served as control,only underwent anesthesia and thoracotomy,without extracorporeal circulation.Group B (n =6) served as underwent deep hypothermic circulatory arrest at 20 ℃ for 120 min,survived for 1 day.Group C (n =6) served as underwent deep hypothermic circulatory arrest at 20 ℃ for 120 min,survived for 2 days.Diffusion-weighted imaging and histopathology were used to study the brain injury.Results There were hematoxylin-eosin (+) and terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (+) cells in cerebral cortex and hippocampus.Hyperintensity was most evident in the cerebral cortex in group B and group C with diffusion-weighted imaging,but it was not seen in hippocampus in both groups.There were 5 and 6 piglets can be seen hyperintensity with diffusion-weighted imaging sequence in group B and group C respectively.Hyperintensity was also seen in T2WI sequence in 3 and 5 piglets in group B and group C respectively.Conclusion The study demonstrates the feasibility of diffusion-weighted imaging on evaluation of brain injury after deep hypothermic circulatory arrest and cerebral cortex was selectively vulnerable to cell injury.It is superior to conventional imaging.

BACKGROUND:In chronic congestive heart failure (CHF),both protein expression and activity of sarcoplasmic reticulum (SR) Ca2+ ATPase (SERCA2a) are decreased, which results in abnormal regulation of calcium ion and systole and diastole dysfunction.OBJECTIVE:To observe the short-and middle-term therapeutic effects of adeno-associated viral gene transfer of SERCA2a in treatment of CHF in rats,and to investigate the possible mechanisms.DESIGN:Randomized-controlled design.SETTING:Department of Geriatric Cardiology and Department of Pathophysiology,General Hospital of Chinese PLA.METHODS:Male Sprague-Dawiey rats,were randomized into 4 groups:sham-operation group,CHF group,CHF+ green fluorescent protein (GFP) group and CHF+SERCA2a group. Gene transfer 10 and 30 days two time points were set in each group.CHF rat models were established by coarctation of abdominal aorta.At postoperative 18 to 20 weeks,the successful rats in CHF group were intraperitoneally injected with 500 μL aseptic normal saline,and those in the CHF+ GFP group and CHF+ SERCA2a group were injected with the same dose of Adeno-associated viral gene with GFP and SERCA2a gene respectively.MAIN OUTCOME MEASURES:①At 10 and 30 days after gene transfer,hemodynamic parameters,the SERCA2a protein expression was analyzed by Western Blot,and SERCA2a activity was carded out by modified Jones method.②The difference of proteome of hearts was detected by expression proteomics in between CHF+ SERCA2a group and CHF group at 30 days.③Electrophoretic separation and quantitation of cardiac myosin heavy chain(MHC) isoforrns were done at 30 days.RESULTS: ①In CHF group and CHF+GFP group, SERCA2a expression and activity were lower than those in sham-operation group.At 10 days after gene transfer.SERCA2a expression was increased by 80.7%and still lower than that of sham-operation group,and activity was also increased by 89.9%,but still lower than that of sham-operation group.And at 30 days,the SERCA2a protein expression and activity in the CHF+SERCA2a group reached to levels of sham-operation group. ②At day 10,LVSP and LV +dp/dtmax and LV-dp/dtmax were increased and LVEDP was decreased dramatically,but they still did not reach to the levels of sham-operation group.At day 30,the parameters were normalized back to control levels.There were no significant differences in each index between CHF+SERCA2a group and sham-operation group.③Overexpression of SERCA2a increased some energy metabolic enzymes of hearts at 30 days.④There were a downregulation of α-MHC and an upregulation of 3-MHC in failing hearts.Overexpression of SERCA2a increased the expression of α-MHC and decreased the expression of β-MHC to the levels of sham-operation group at 30 days.CONCLUSION: Adeno-associated viral gene transfer of SERCA2a can enhance SERCA2a functions, maintain calcium homeostasis,improve cardiac energy metabolism,and normalize the expression of MHC isoforms in CHF rats.As a result,the heart functions can be improved. So adeno-associated viral gene transfer of SERCA2a has good therapy effects on CHF rats in short and relatively long periods.

Objective To investigate the G protein-coupled receptor kinase 2 (GRK 2) level in peripheral blood lymphocytes with cardiac func-tion in elderly patients with acute myocardial infarction. Methods This study enrolled 40 patients with acute ST-segment elevation myo-cardial infarction (STEMI) and 40 patients with unstable angina. All patients were 65 years or older. Cardiac function was evaluated by echocardiography, and the GRK 2 level in peripheral blood lymphocytes was measured. Patients with STEMI were followed up for 2 years. Results The GRK 2 level in peripheral blood lymphocytes was significantly higher in patients with STEMI than in patients with unstable angina, and was negatively correlated with left ventricular ejection fraction, cardiac output, stroke volume, and left ventricular fractional shortening. The GRK 2 level was significantly elevated in some patients with acute STEMI and poor cardiac function. Conclusions In-creased GRK 2 level in patients with acute STEMI may contribute to poor myocardial systolic function and myocardial remodeling. Meas-urement of the GRK 2 level in peripheral blood lymphocytes may assist in the evaluation of cardiac function and myocardial remodeling in elderly patients with acute STEMI.

Objective To analyze the characteristic of trauma in 246 fatal falls. Methods We collect 246 cases of fatal falls from Pudong district of Shanghai from 2005 to 2014, and then analyze the characteristics of trauma by using different parameters. Results The height in suicides by fall is higher than accidents by fall (25.7m:13.98m). In all fatal falls, head has a higher possibility in suffering trauma than other body regions (75.2%), while neck has a lower possibility (6.5%). The possibility of head trauma in accidents by fall is higher than suicides by fall (89.4%:75.2%). Conclusion Comparing with suicides by fall, victims in accidents by fall have lack of protection from limbs. Besides, the low rate of ifnding neck trauma suggests legal physicians should be more careful in examining this area.

Objective To compare the clinical features between radiological isolated syndrome (RIS) and classical multiple sclerosis (CMS), in order to improve the understanding of the RIS. Methods All 35 patients with RIS and 32 patients with CMS were selected. The epidemiological and clinical findings, cerebrospinal fluid, neural electrophysiological examination and magnetic resonance imaging (MRI) data were analyzed. Results There were no statistical differences in sex ratio and onset age between RIS patients and CMS patients (P>0.05). The main symptoms of in patients with RIS were headache (45.7%, 16/35), dizziness (40.0%, 14/35), hypomnesis (20.0%, 7/35) and psychiatric disorders (11.4%, 4/35). But the main symptoms of in patients with CMS were limb weakness (75.0%, 24/32), sensory abnormalities (68.8%, 22/32) and ocular symptoms (34.4%,11/32). The incidences of limb weakness, sensory abnormalities and ocular symptoms in patients with CMS were significantly higher than those in patients with RIS:75.0%(24/32) vs. 0, 68.8%(22/32) vs. 0 and 34.4%(11/32) vs. 0, and there were statistical differences (P<0.01). The 18 patients with RIS and 21 patients with CMS underwent the examination of cerebrospinal fluid, and there was no significant difference in leukocyte between patients with RIS and patients with CMS (P>0.05). The cerebrospinal fluid protein and the incidences of IgG index>0.7 in patients with RIS were significantly lower than those in patients with CMS:0.175 (0.03-0.69) g/L vs. 0.440 (0.04-1.09) g/L and 3/18 vs. 47.6%(10/21), and there were statistical differences (P<0.05). The 15 patients with RIS and 22 patients with CMS underwent the examination of neural electrophysiological, and the abnormality rates of visual evoked potential (VEP) and brain stem auditory evoked potential (BAEP) in patients with RIS were significantly lower than those in patients with CMS:4/15 vs. 63.6%(14/22) and 3/15 vs. 54.5%(12/22), and there were statistical differences (P<0.05). But there was no statistical difference in the abnormality rate of somatosensory evoked potential (SEP) between patients with RIS and patients with CMS (P>0.05). On MRI, the demyelinating lesions of RIS and CMS were both mainly distributed in the periventricular, semi-oval center, infratentorial white matter, partly involving corpus callosum or cortical. The rates of demyelinating lesions in brainstem and cerebellum in patients with RIS were significantly lower than those in patients with CMS:5.7%(2/35) vs. 34.4% (11/32) and 2.9% (1/35) vs. 25.0% (8/32), and there were statistical differences (P<0.01 or <0.05). Comparison with CMS lesions, RIS lesions mainly showed patching and stippled, and there were statistical differences (P<0.01 or <0.05). The rates of lesions enhancement and spinal cord injury in patients with RIS were significantly lower than those in patients with CMS: 2/17 vs. 45.0% (9/20) and 1/14 vs. 43.5% (10/23), and there were statistical differences (P<0.05). Conclusions There are differences in clinical findings, cerebrospinal fluid, neural electrophysiological examination and MRI appearances between RIS and CMS.

Objective To establish three methods of DNA extraction from Cryptosporidium parvum oocysts and test by PCR. Methods After three freeze-thaw cycles, three kinds of templates were extracted from the oocysts by Chelex-100, phenol/chloroform or genomic DNA purification system kit, and used for PCR detection. According to the sequence of a C.parvum gene (L16996), a pair of primers was designed and synthesized, and used for PCR. The sensitivity of the template by Chelex-100 method was also tested by PCR. Results One 446 bp PCR product was observed by agarose gel electrophoresis for all three kinds of templates. The PCR sensitivity by Chelex-100 extracted DNA reached for detection of a specimen containing only 1/2 oocyst. Conclusion The three kinds of extraction can all be served as templates for PCR detection of C.parvum oocysts, while Chelex-100 method is simpler, quicker and more reliable for DNA extraction of the parasite.