To investigate the HBV quasispecies groups in the patients with chronic HBV infection. A set of specific primers was synthesized according to HBV DNA sequence of the Chinese strain, and the whole pre core/core region was amplified by PCR method from the serum of 4 patients with chronic HBV infection.Then the PCR products were subcloned into pGEM Teasy vectors. Clones were selected to be sequenced,and then sequence comparison was made to find the difference. Each sequence of selected clones was found to be different. The homology of the nucleic acid sequence from the same patient was above 98 0%. There were many mutation types in this region, including point substitution, core region internal deletion, as well as frame shift reading. The results indicated that there were HBV quasispecies groups in chronically infected patients. The mutation in this region may influence the prognosis of HBV infection.

T7 cDNA phage display method was employed to find the binding protein of PreSl protein. PreSl protein was coated in a 96-well ELISA plate, and then T7 cDNA library phages were bound to the target protein. Phages which did not bind to garget protein were washed away and the binding phages were eluted. Insertions from different clones were sequenced, and the deduced amino acid sequences were analyzed by Vector 6.0 software. Using BLAST software in GenBank, whole length of amino acid sequence of binding protein was obtained. After 4 rounds of biopanning, recombinant T7 phages with binding ablity were amplifed by infection to E. coli. One piece of amino acid sequence was found to be amino terminal of product of glioma tumor suppressor candidate region gene 2 (GLTSCR2). There was a binding domain KxPxKSGxxxL in these clones. T7 cDNA phage display technique can be used bo find the ligand. GLTSCR2 coding protein may be the binding protein to preSl protein of HBV.\;

Objective To study whether abnormalities can be detected by MR diffusion tensor imaging (DTI) technology in radiologically isolated syndrome (RIS) patients with normal-appearing white matter (NAWM).Methods Twenty-seven patients who met diagnostic criteria for RIS were collected.Sixteen age-and sex-matched healthy controls with normal neurologic examination findings and no history of neurologic or psychopathic disorders were included.All subjects were examined by both conventional scan and DTI scan on GE Signa 3.0 T MRI.All the images were transmitted to the Advantage Workstation 4.2P and postprocessed using functool software.Anisotropic maps and average diffusion coefficient (ADC) maps were reconstructed.Two ROIs were selected in the genu and splenium of the corpus callosum separately at the trigone of lateral ventricle level.Other 2 ROIs were selected in the anterior and posterior body of the corpus callosum separately at the level of the lateral ventricle body,and 1 ROI was selected in the white matter of bilateral frontal and occipital lobe separately,then the FA and MD values were measured.The diffusion indices (FA and MD)were analysed by SPSS 13.0.Independent-sample t test was performed to examine the group differences in each subregion.Intraclass correlation coefficient analysis was performed to assess the diffusion indices of two measurements in each subregion.Results The ICC of 2 measurements was 0.934-0.989 (P ＜ 0.01),which showed favorable consistency.The FA values were decreased obviously in the genu,anterior and posterior body of the corpus callosum of the RIS patients compared with controls (0.705 ±0.040 vs 0.738 ±0.045,0.632 ±0.043 vs 0.675 ±0.042,0.628 ±0.043 vs 0.666 ± 0.045,t =-3.526,-4.487,-3.890,P ＜0.01),but the FA values of the two groups did not show any significant difference in the splenium,the white matter of the frontal and occipital lobe respectively (t =-1.387,-0.683,-1.243,P ＞0.05).In comparison with controls,the RIS patients had increased MD values in the genu,anterior and posterior body of the corpus callosum.(0.891 ±0.038 vs 0.874 ±0.035,0.839 ± 0.047 vs 0.794 ± 0.031,0.833 ± 0.039 vs 0.792 ± 0.057,t =2.101,5.836,5.146,P ＜ 0.05),but the MD values of the two groups did not show any significant difference in the splenium,the white matter of the frontal and occipital lobe respectively (t =1.671,1.702,1.624,P ＞ 0.05).Conclusion The NAWM abnormalities in the patients with radiologically isolated syndrome could be detected by DTI.

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 explore any changes in the surface electromyography (sEMG) signals measured on the spastic upper limb muscles of stroke parents during maximum isometric voluntary contraction and to analyze any abnormal synergy patterns quantitatively in order to design better rehabilitation programs for developing coordination.Methods Ten stroke survivors with hemiparesis were selected into a patient group and ten healthy counterparts were recruited into a control group.sEMG signals were recorded bilaterally from the flexor carpi ulnaris (FCU),biceps brachii (BB),triceps brachii (TB) and deltoid (D) during maximum isometric voluntary contractions involving wrist flexion and extension,elbow flexion and extension,and shoulder abduction.The two groups' co-contraction ratios (CR) and co-activation ratios were calculated and compared.Results During elbow flexion and extension the average CR of the BB on the affected side was significantly higher than that on the unaffected side and also significantly higher than the control group average.The average CR of the TB on the affected side was significantly higher than that of the healthy controls.In all cases the average CR of the BB was larger than that of the TB.The difference in CR between the TB and the BB on the affected side was significantly larger than on the unaffected side and the control group average.During elbow flexion,the co-activation ratio of the FCU,TB and D on the affected side was significantly higher than on the unaffected side and among the healthy controls,and the co-activation ratio of the FCU on the affected side was significantly higher than that of the D and TB.During elbow extension,the co-activation ratio of the FCU,BB and D on the affected side was significantly higher in the same way,and the co-activation ratio of the FCU on the affected side was again significantly higher than that of the D and BB.During wrist flexion,the average co-activation ratio of the BB and D on the affected side was significantly greater than that on the unaffected side and among the healthy controls,and the co-activation ratio of the BB on the affected side was significantly higher than that of the D and TB.During shoulder abduction,the co-activation ratio of the BB on the affected side was significantly larger than on the unaffected side and among the healthy controls.Conclusion After a stroke the upper limbs often show flexor spasticity and abnormal synergy patterns.Rehabilitation strategy should be adopted to tackle these so as to enhance overall limb coordination.

Objective:To investigate the effects of 125I radioactive particle implantation in the treatment of maxillofacial malignancy. Methods:43 patients with maxillofacial malignancy were treated with 125I radioactive particle implantation.The procedure was carried out according to the treatment planning system(TPS),with the particles spaced uniformly at 1 cm intervals and with the activity of 0.7 m Ciby 41 particles per case on a verage.All patients were followed up for 6-60 months.Results:The whole treatment procedurewas successful,and no particle displaced.The follow-up rate was 93.02% and treatment effective rate(CR+PR) was 90.70%.Norecur-rence was foundinall target areas.The mortality due to tumor was 9.30%and total survival rate was 74.43%.The cumulative survival rate of the patients in 0.5,1,2,3 and 5 years was 93.0%,85.7%,79.3,69.8% and 56.9% respectively.Survival periodon aver-age was 36.06-50.04 months,with the median of 43.05 months.The longest tumor-free survival period was 60 months.Radiation in-jury rate was 20.93% and only level 1 radiation injury was observedinall the cases.Facial nerve dys function was found in 2 cases and recovered after treatment.Conclusion:Treatment of maxillofacial malignancy by implantation of 125I particles is convenient and mini-mally invasive.The treatment canincrease survival rate of the patients and guaran tee the oropharynxes' function.

Objective To purify pre-ribosome and ribosome of mammalian ceils using continuous sucrose density gradient ultracentrifugation.Methods Continuous sucrose density gradient was established by ultracentrifugation,and the continuous sucrose density gradient of 10％-30％ and 10％-45％ were used to extract the pre-ribosome and ribosome in mammalian cells,respectively.The mammalian cell lysis buffer was added to the established continuous sucrose density gradient.Pre-ribosome and ribosome with different sedimentation coefficients were collected and the A260 absorbance of each sample was measured.Proteins of each sample were extracted to detect the large subunit protein,RPL15 by Western Blot.Results Large subunit ribosomal protein RPL15 exists on 60S of the pre-ribosome,and also on 60S,80S and polyribosome of mature ribosome.Conclusions The continuous sucrose density gradient,which is established by the swing-out rotor,can be used to isolate the pre-ribosome and ribosome of mammalian cells rapidly.This method has the advantages of good separation effect and simple operation,which provides a good method for rapid and large amount preparation and separation of various kinds of ribosomes.

Objective To investigate the process that chromosome kinesin KIF4A promote cisplatin resistance in lung cancer cells.Methods Reverse transcription PCR (RT-PCR) and Western Blot experiments were performed to analyze the expression of KIF4A in lung cancer cells A549 and cisplatin (DDP) resistant cells A549/ DDP.Cell transfection, RNA interference (RNAi) experiments and thiazolyl blue tetrazolium bromide (MTT) assays were carried out to examine cell proliferation of A549 cells with overexpression of exogenous KIF4A and A549/DDP cells with depletion of endogenous KIF4A after cisplatin treatment.Results Expression of KIF4A in A549/DDP cells was higher than that in A549 cells.With overexpression of exogenous KIF4A, A549 cells displayed drug resistance to cisplatin.On the contrary, depletion of endogenous KIF4A in A549/DDP cells resulted in cisplatin sensitivity.Conclusions Chromosome kinesin KIF4A involves in the regulation of cisplatin resistance in lung cancer cells and KIF4A may be a potential and effective new biological target for treatment of lung cancer cisplatin resistance.

Objective:To analyze the clinical characteristics of patients with femoral neck fracture in Tianjin Hospital.Methods:Data of femoral neck fracture patients discharged from Tianjin Hospital from January 1, 2016 to December 31, 2017 were retrospectively analyzed. The gender, age, fracture type, injury mechanism and treatment method were collected.Results:Total of 2,150 eligible patients including 736 males and 1414 females were selected. There were significant differences in the gender distribution in different age groups ( χ2=91.63, P<0.001). Among young patients, males were more than females, while among middle-aged and elderly patients, females were more than males. The main type of fracture was displaced type (75.56%, 1,646/2,150), the displaced rate was the lowest in young patients (55.17%, 64/116), while it was the highest in old patients (81.91%, 1,159/1,415). The main injury mechanism of the three age groups were falls [young 61.21% (71/116), middle 80.29% (497/619), eldly 91.24% (1,291/1,415)]. There was a significant difference in the treatment methods between different age groups ( χ2=1,057.11, P<0.001). The main treatment method for young and middle-aged patients was internal fixation, no matter whether the fracture was displaced or not. The proportion of undisplaced patients with internal fixation (86.22%, 169/196) was higher than that of displaced patients with internal fixation (62.88%, 266/423) in middle-aged group ( χ2=34.93, P<0.001). In the elderly, more patients used internal fixation in undisplaced group, while more patients used hemiarthroplasty in displaced group. The age of old femoral neck fracture (median age was 74) was older than that of fresh fracture (median age was 70) ( Z=-2.777, P=0.005). And the displaced rate of patients with old femoral neck fracture (92.41%, 73/79) was higher than that of the fresh ones (75.95%, 1,573/2,071)( χ2=11.48, P=0.001). The patients with old femoral neck fracture usually adopt total hip replacement, while the proportion of three kinds of operation (internal fixation, hemiarthroplasty, total hip replacement) were similar among the fresh ones, and the internal fixation was the most. Conclusion:Femoral neck fracture is the most common in elderly women. The patients with displaced fracture are more than that with undisplaced fracture. Falling is the main injury mechanism in both young and old people. Internal fixation is commonly used in young and middle-aged patients, while arthroplasty is often used in elderly patients.

Objective To establisha multiplex STR genotyping m ethod for autosom al STR and Y-STR loci in forensic biological practice. Methods W idely used autosom al STR loci and Y-STR loci w ere se-lected. A set of PC R prim ers w as designed, and a 5-dye fluorescent labeled STR multiplex PC R reagent kit w as developed. Results A kit w as developed w hich can sim ultaneously detect 15 autosom al STR loci, 10 Y-STR loci, and an Amelogenin. Conclusion The 15 autosom al STR plus 10 Y-STR kit in com bination w ith capillary electrophoresis m ethod w as used to STR genotyping w ith accurate and reli-able results. The new one-step testing kit can potentially be w idely used in forensic cases and D N A databank in the future.