Objective To generate HLA - A * 0201 and A * 2403 restricted HBcAg - specific cytotoxic T lymphocyte (CTL) clones. Methods Peripheral blood mononuclear cells (PBMCs) were derived from a HLA - A * 0201/2403 - positive patient with chron- ic hepatitis B . PBMCs were stimulated respectively using two synthetic peptides( HBc18 ～ 27 and HBc 117 ～ 125) , and epitope -specific CTL clones were generated by limiting dilution technique with PHA alone or combined with synthetic peptides. The cell clones were then characterized by IF staining,FCM and LDH release. Results After 2 weeks of in vitro stimulating PBMCs, specific CTL lines were established. 29 T clones were generated from those CTL lines using HBc18 ～27 stimulating. Among the 29 clones ,28 clones belonged to CD8~+ T cells and all displayed cytolytic activity. 12 CD8 +T clones were generated from those CTL lines using HBc117 ～ 125 stimulating. Specific cytotoxic activity was observed in 9 of those clones. The other three displayed less cytolytic activity. In the process of cloning, PHA was used alone or combined with synthetic peptides,and the achievement showed a rate of 15.62% and 14.58% . Conclusion HBc18 ～ 27 and HBc117 ～ 125 are capable of activating CD8~+ T cells in PBMCs of HLA - A * 0201 /2403 patient with chronic hepatitis B. In the process of CD8~+T cloning, synthetic peptides could not increase the rate of successful cloning.

BACKGROUND:Cerebral ischemia tolerance can promote proliferation of autologous neural stem cells in the hippocampus of cerebral infarction rats, but panaxtrial saponins effects on the proliferation of autologous neural stem cells in the brain have not been reported. OBJECTIVE:To explore the relationship of panaxtrial saponins, ischemic preconditioning and proliferation of endogenous neural stem cells in the hippocampus of rats at 7 days after cerebral infarction, and to observe the effect on neurobehavioral scores of rats after cerebral infarction. METHODS:Fifty Sprague-Dawley rats were included and randomly divided into five groups:sham group, ischemia group, ischemic control group, ischemic preconditioning group, and panaxtrial saponins group. In the latter four groups, acute models of cerebral infarction were established using Zea-Longa method. In the sham group, only an incision was made on the neck. The focal-focal ischemic tolerance models were established with twice suture method in the ischemic preconditioning and panaxtrial saponins groups. Sham operation was instead of ischemic preconditioning in the ischemic control group. In the panaxtrial saponins group, rats were given intraperitoneal injection of 100 mg/kg panaxtrial saponins at 7 days before modeling. RESULTS AND CONCLUSION:After 7 days of cerebral infarction, the neurobehavioral score and the number of neural stem cells in the hippocampus were significantly increased in the ischemia group (P<0.01);compared with the ischemia group, the neurobehavioral scores were lowered in the ischemic preconditioning and panaxtrial saponins groups (P<0.01), while the number of neural stem cells in the hippocampus was increased (P<0.01). However, there was no difference between the ischemic preconditioning and panaxtrial saponins groups (P>0.05). In addition, differences in the neurobehavioral scores and the number of neural stem cells in the hippocampus were insignificant between the ischemic control group and ischemia group (P>0.05). These findings indicate that panaxtrial saponins can play a role similar to ischemic tolerance, and thus improve neurologic impairment in rats with cerebral infarction.

Objective To investigate the effect of brain ischemic preconditioning (IP) combined with traditional Chinese medicine three seven three alcohol saponin (PTS) on proliferation of endogenous neural stem cells and the mRNA expressions of delta opioid receptor (DOR),Bax,Bcl-2 in hippocampus at 7d post middle cerebral artery occlusion (MCAO).Methods The focal-focal ischemic tolerance models were established with twice suture method.80 SD rats were included and randomly divided into 5 groups:sham group,MCAO group,sham+ MCAO group,IP+ MCAO group,PTS+MCAO group (n=16 each).We chose 10 SD rats from each group to evaluate their neurological status,and made BrdU fluorescent immunolabeling.In addition,we chose the other 6 SD rats to detect the expression levels of DOR,Bax and Bcl-2 mRNA in ischemic region in hippocampusby using RT-PCR.Animals were given one set of BrdU injections (on day 6,three times,4h apart,50mg/kg) to label the proliferating cells.The neurological status was assessed by using Zea Longa neurological deficit scores at 7 days following cerebral infarction.Results Zea longa neurologic deficit scores in MCAO group and sham+ MCAO) group had significantly differences with IP+ MCAO group and PTS+ MCAO group respectively at 7d post MCAO(P＜0.01).There was no significant differeuce in Zea-longa neurologic deficit scores between MCAO group versus sham+ MCAO group,and IP+ MCAO group versus PTS+ MCAO group(P＞0.05).The number of BrdU+ ceils in hippocampus had significant differences between IP+ MCAO and PTS+ MCAO groups at 7d post MCAOand three groups of sham,MCAO and sham+ MCAO respectively (P＜0.01).There was no difference in the number of BrdU+ cells between MCAO versus Sham + MCAO groups and IP + MCAO versus PTS+MCAO groups(P＞0.05).DOR and Bcl-2 mRNA expression levels were higher and Bax mRNA expression level was lower in IP+ MCAO group than in MCAO,Sham+ MCAO and PTS+MCAO groups (P＜0.01).There were no significant differences in DOR,Bcl-2 and Bax mRNA expressions among MCAO,Sham + MCAO and PTS + MCAO groups (P＞ 0.05).Conclusions Acute cerebral infarction can induce the proliferation of endogenous neural stem cells in hippocampus in SD rats.IPC can facilitate the proliferation of endogenous neural stem cells in hippocampus afteracute cerebral infarction,improve the symptoms of neurologic dysfunction,increase DOR and Bcl 2 mRNA expressions,and reduce Bax mRNA expression in SD rats.PTS can facilitate the proliferation of endogenous neural stem cells in hippocampus after acute cerebral infarction in SD rats,and improve the symptoms of neurologic dysfunction,but it has no influence on the expressions of DOR,Bcl-2 and Bax mRNA.

Objective To explore the effect of δ-opioid receptor (DOR)on ischemic tolerance of rat brain.Methods The focal ischemic tolerance models of Sprague Dawley rats were established using the twice suture method with the middle cerebral artery occlusion (MCAO).A total of 24 male Sprague Dawley rats were randomly divided into sham group,ischemia (MCAO) group,sham +ischemia (sham+ MCAO) group and ischemic preconditioning + ischemia (IP+ MCAO) group (n=6,each).The neurological status was assessed using Zea-Longa neurological deficit scores at 7 days after cerebral infarction.The mRNA expressions of DOR,Bax,Bcl-2 in hippocampus in ischemic rat brain were detected by RT-PCR method.Results The Zea-Longa neurological deficit scores were 0.0±0.0,2.6±0.5,2.8±0.6 and 1.5±0.6 in Sham group,MCAO group,Sham+MCAO group and IP+ MCAO group,respectively at 7 days after cerebral infarction.The scores had significant differences among MCAO group,Sham+ MCAO group and IP+ MCAO group (both P＜0.01),but no difference between MCAO group and Sham+MCAO group(P＞0.05).The mRNA expressions of DOR and Bcl-2 were higher and Bax mRNA expression was lower in IP+MCAO group than in MCAO and Sham+ MCAO groups (all P＜0.01).Conclusions Ischemic preconditioning may increase the mRNA expressions of DOR and Bcl-2,reduce Bax mRNA expression,and improve the neurological status in rats.

Objective To aim at the problem of bedsore which is always occurred on the crowd of the aging population,empty-nest family and the disabled patients etc,a two-way side turn over mechanism for intelligent sanitation nursing instrument is designed.Methods The turn over angle of the mechanism was analyzed and calculated after establishing the mechanism's three-dimensional model.The forces situation of the state of lying,side lean against and left/right side turn over were carried out through finite element analysis by using of Pro/Mechanica.Results The results showed that the sum of two-way side turn over mechanism's two-way side turn over angle was 30 degrees.The materials met the mechanism's force requirement.Conclusions The results can provide a theoretical basis for the designer to determine the structure parameters and sizes of mechanism.

Objective To investigate the effect of focal cerebral ischemic preconditioning (IPC) on brain edema and the expression of nuclear factor-?B( NF-?B) and its target gene MMP-9. Methods Forty-five SD rats were divided into 3 groups in which control group received sham surgery only, and the other two groups received 2 hours of middle cerebral artery occlusion (MCAO) followed by 22 hours of reperfusion with or without 10 minutes of IPC 3 days before. Brain water content, expression of NF-?B and MMP-9 mRNA were evaluated in each group by wet-dry weight method, immunohistochemistry staining and RT-PCR. Results Compared with the SS group, there was a lower NF-?B immunoreactivity and MMP-9 mRNA level (16 098.2?1 265.3 vs 23 565.8?1 978.4,50.7% vs 84.1%, P

Starting from the brand new angle of exploring the interaction between the knowledge capital and the financial capital of the hospital, the paper presents the theoretical basis and practical significance of the interaction between the knowledge capital and the financial capital of the hospital. Moreover, by establishing tentative models of hospital capital operation and by making theoretical analyses into the interactive and incremental relationship between knowledge capital and financial capital by means of mathematical analysis and deduction, the paper further demonstrates that the interactive integration of the knowledge capital and the financial capital of the hospital is an important condition which ensures that the operation of the hospital will maintain a high momentum and its comprehensive competitiveness will continue to be in a power position.

Objective To observe the effect of focal cerebral ischemic preconditioning on the expression of glial fibrillary acidic protein (GFAP) and to investigate the significance of astrocyte activation in cerebral ischemic tolerance.Methods Thirty-six healthy male SpragueDawley rats were randomly divided into reischenmic,ischemic and control groups (n = 12 in each group) after ischemic preconditioning.The former two groups received 10 minutes middle cerebral artery occlusion (MCAO) preconditioning or sham operation 3 days before the 2-hour MCAO.The rats were killed 24 hours after the second MCAO.The control group only receivedthe two sham operations with an interval of three days.The infarct volume,histopathological changes,and GFAP expression in each group were compared.Results The infarct volume after ischemic preconditioning in the reischenmic and ischemic groups was 136.85 ± 14.51 mm3and 281.37 ± 29.93 mm3 respectively.The former was significantly reduced 53.15%compared to the latter (P =0.007).At the same time,neuronal degeneration and necrosis was reduced significantly,and GFAP expression was upregulated significantly (the mean absorbance for immunohistochemical staining in both groups was 102.66 ± 8.39 and 86.28 ± 6.19respectively,P = 0.009) after ischemic preconditioning in the reischemic group.Conclusions Focal ischemic preconditioning may induce brain ischemic tolerance and promote GFAP expression.The activation of astrocytes may be one of the mechanisms of cerebral ischemic tolerance.

Objective To evaluate the effect of endoscopic management of foreign bodies in the upper digestive tract. Methods Clinical data and endoscopic treatment methods of 41 patients were retrospectively analyzed from October 2014 to May 2016. Patients with incomplete medical records were excluded. Results Foreign bodies in the upper digestive tract occurred high frequency in elderly. 53.6% of the foreign bodies were located in the esophagus. Date stones was the main type of foreign bodies (56.1%). 41 cases with foreign bodies in digestive tract were successfully extracted, while 1 case occurred perforation. Conclusion Endoscopic management of gastrointestinal foreign bodies is safe and effective.

Objective:To establish a fluorescent reporter human induced pluripotent stem cell line (hiPSCs) for monitoring the expression of visual system homeobox 2 ( VSX2). Methods:VSX2_small guide RNA (sgRNA) was inserted into vector PX459 to construct knockout plasmid, and the P2A-eGFP knock-in donor plasmid was conducted at the same time.The two plasmids were transfected into BC1-hiPSCs.Single cell clones were generated after treatment of puromycin.Correct insertion was confirmed by PCR and Sanger sequencing.The isogenicity of the parental and the reporter hiPSCs was confirmed by STR analysis and karyotyping.Pluripotency capacity of the reporter hiPSCs was analysed by reverse trascription PCR and immunofluorescence.Three-germ-layer formation experiment was carried out to analyse the multi-lineage differentiation ability of the reporter hiPSCs.The reporter hiPSCs were further differentiated to obtain three-dimension (3D) retinal organoids, and immunofluorescence was used to identify the co-localization of the enhanced green fluorescent protein (eGFP) and VSX2.Results:A VSX2 eGFP reporter hiPSC clone was successfully obtained by CRISPR/Cas9 technology, which was consistent with the parental hiPSCs (BC1-hiPSCs) in morphology, without any chromosomal aberrations or cell line cross-contamination.Reverse transcription PCR assay and immunofluorescence showed obvious positive expressions of iPSCs markers in BC1- VSX2 eGFP-iPSCs, including NANOG, OCT4, SOX2, DNMT3B and GDF3 mRNA as well as NANOG, OCT4, SSEA4 and TRA-1-60 protein.The α-fetoprotein (AFP), α-smooth muscle actin (α-SMA) and neuronal class Ⅲ β-tubulin (TUJ1) were expressed in endoderm, mesoderm and ectoderm, respeetively, derived from BC1- VSX2 eGFP-iPSCs, and eGFP and VSX2 were co-stained in the neural retinal layer of 3D retinal organoids derived from BC1- VSX2 eGFP-iPSCs by immunofluorescence. Conclusions:VSX2 fluorescent reporter hiPSCs is successfully generated, which can monitor the temporal and spatial expression changes of VSX2 protein in real time, providing a powerful tool for evaluation of retina development mechanism and cell therapy.