Objective:To develop a catalytic system for the asymmetric Morita-Baylis-Hillman ( MBH) reaction of conjugated ni-troalkene with activated aldehyde, and screen out the chiral catalysts with high activity and enantioselectivity. Methods: Totally 21 chiral organocatalysts were applied in the asymmetric MBH reaction ofβ-nitrostyrene with ethyl glyoxylate, and the ee value was deter-mined by chiral HPLC. The effects of temperature, solvent and substrate ratio on the catalytic reaction were investigated. Results: In the presence of cinchona alkaloid catalyst (DHQ)2AQN, β-nitrostyrene reacted with ethyl glyoxylate in toluene at 0℃ affording the MBH adduct in 60% yield with good enantioselectivity (up to 56.9% ee). Conclusion: The bis-cinchona alkaloids with aromatic bridging group are the efficient catalysts for the asymmetric MBH reaction ofβ-nitrostyrene with ethyl glyoxylate, and moderate isolated yield and enantioselectivity are obtained.

Objective To evaluate the effect of hydrogen-rich saline on the expression of phosphor-p38 mitogen-activated protein kinase (p-p38MAPK) during cerebral ischemia-reperfusion (I/R) in rats.Methods Seventy-two adult male Sprague-Dawley rats,weighing 220-250 g,were randomly divided into 3 groups (n =20 each) using a random number table:sham operation group (group S),I/R group and hydrogen-rich saline group (group I/RH).Cerebral ischemia was induced in chloral hydrate-anesthetized rats by 2 h middle cerebral artery occlusion in I/R and I/RH groups.The artery was only exposed but not occluded in group S.At 3 days before operation and immediately after onset of reperfusion,hydrogen-rich saline (0.6 mmol/L) 10 ml/kg was intraperitoneally injected in group I/RH,while the equal volume of normal saline was given in S and I/R groups.Neurological deficits were blindly assessed and scored at the end of 24 h reperfusion.The animals were then sacrificed,and brains were removed for microscopic examination and for determination of the cerebral infarct size (by TTC),brain water content,cell apoptosis (by TUNEL),and expression of p38MAPk and phosphor-p38MAPK (p-p38MAPK) (by immunohistochemistry and Western blot).Apoptosis index was calculated.Results Compared with group S,neurological deficit score,apoptosis index,brain water content and cerebral infarct size were significantly increased,and the expression of p38MAPK and p-p38MAPK was up-regulated in I/R and I/RH groups.Compared with group I/R,neurological deficit score,apoptosis index,brain water content and cerebral infarct size were significantly decreased,and the expression of p38MAPK and p-p38MAPK was down-regulated in group I/RH.The pathological changes of cerebral tissues were significantly attenuated in group I/RH as compared with group I/R.Conclusion Hydrogen-rich saline can reduce cell apoptosis through inhibiting p-p38MAPK expression,thus attenuating cerebral I/R injury in rats.

Objective To evaluate the effect of hydrogen on the activation of caspase-3 in brain tissues during cerebral ischemia-reperfusion (I/R) in rats.Methods Thirty-six healthy adult male Sprague-Dawley rats,weighing 220-250 g,were divided into 3 groups (n=12 each) using a random number table:sham operation (group S),I/R group and hydrogen group (group H).Cerebral ischemia was induced by occlusion of the middle cerebral artery followed by reperfusion in I/R and H groups.In group H,hydrogen-rich saline 5 ml/kg (0.6 mmol/L) was injected intraperitoneally at 3 days before establishment of the model and immediately after the onset of reperfusion.At 24 h of reperfusion,the rats were sacrificed,and hippocampal tissues were obtained for determination of neuroapoptosis (by TUNEL),apoptotic neuron count and expression of activated caspase-3 (by Western blot).The brain tissues in the ischemic area were obtained and stained with haematoxylin and eosin for examination of the pathological changes.Results Compared with group S,the expression of activated caspase-3 was significantly up-regulated,and the apoptotic neuron count was increased in I/R and H groups (P<0.05).Compared with group I/R,the expression of activated caspase-3 was significantly down-regulated,the apoptotic neuron count was decreased (P<0.05),and the pathological changes of brain tissues were significantly reduced in group H.Conclusion The mechanism by which hydrogen inhibits neuroapoptosis during cerebral I/R is probably related to inhibited activation of caspase-3 in brain tissues of rats.

HIV-1 integrase (IN) is a key enzyme for the viral replication. The protein-protein interaction (PPI) between HIV-1 IN and a cellular cofactor lens epithelium-derived growth factor (LEDGF/p75) is a validated target for anti-HIV drug discovery. In order to build the platform for screening inhibitor against PPI between IN and LEDGF/p75, the vector containing the LEDGF/p75 protein cDNA was constructed and expressed in Escherichia coli and the function of the LEDGF/p75 protein was assayed. The LGDGF/p75 encoding gene optimized according to the preference codon usage of E. coli, was synthesized and cloned into the expression vector pGEX-4T-1 to form a recombined plasmid, then transformed into host cell E. coli BL21 (DE3). The recombined clones were identified and confirmed by BamH I/Sal I digestion and sequencing, the successfully recombined plasmid in the host cell was induced by IPTG and the condition of the expression was optimized. The expressed protein was purified by the Ni2+ affinity chromatography column and SDS-PAGE was used to analyze the molecular weight and specificity. In addition, ELISA assay was used to analyze the function of the recombinant protein. The recombinant LGDGF/p75 was soluble, and expressed highly and stably in E. coli. The protein was proved to enhance HIV-1 IN strand transfer activity in vitro by ELISA. It will be helpful to build the platform of screening inhibitors against PPI between IN and LEDGF/p75.
Cloning, Molecular ; Escherichia coli ; metabolism ; HIV Integrase ; metabolism ; HIV-1 ; physiology ; Humans ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; Protein Binding ; Virus Replication

The cervical sagittal balance is a description of cervical spine at the standing position. The position of cervical spine is measured by the corresponding sagittal parameters. The commonly used sagittal parameters of cervical spine include cervical lordosis (CL), cervical sagittal vertical axis (cSVA), T 1 Slope (T 1S), etc. Cervical sagittal balance is closely related to the overall balance of the spine, and the imbalance of sagittal parameter is also related to the progression of cervical disease, the choice of surgical methods and the postoperative efficacy. In recent years, more and more researchers have begun to pay attention to the sagittal balance of the cervical spine, especially the variation of sagittal parameters after cervical surgery, which can predict the postoperative neurological recovery and the change of long-term efficacy or complications. However, due to the variety of cervical surgery methods and the variety of cervical sagittal balance parameters, the existing studies can only reflect the relationship between the two from a certain aspect, while lacking overall and comprehensive description. In this paper, the relationship between sagittal plane balance and cervical surgery was reviewed by systematically searching literatures and screening relevant studies. According to our review, the cervical sagittal parameters have different tendency after different cervical surgery. Generally, anterior cervical surgery can maintain or improve the sagittal plane balance of the cervical spine, among which anterior cervical discectomy and fusion (ACDF) surgery has a best effect.After posterior cervical surgery, the cervical sagittal parameters tend to be unbalanced or compensated, and patients with preoperative sagittal imbalance, advanced age, and complex OPLL are more likely to have postoperative imbalance or imbalance aggravation. Therefore, attention should be paid to preoperative evaluation and reasonable surgical design in clinical practice to prevent the occurrence of sagittal imbalance of patients after surgery.

Based on the infrared spectra of Lophatheri Herba and Commelinae Herba, one-dimensional infrared spectra, second derivative spectra and two-dimensional correlated spectra were used to find out the differences between Lophatheri Herba and its imitations, respectively. The common peak ratio and variant peak ratio dual-indexes sequential were calculated and established according to infrared spectra of eleven batches of herbs. Infrared spectral data of Lophatheri Herba cluster analysis was applied to explore the similarity between each sample. The grouping results trend of sequential analysis of dual-indexes and cluster analysis was accordant. The results showed that the differences could be found by multi-level identification, and the source and the quality of the herbs could be effectively distinguished by the two analysis methods. Infrared spectroscopy, used in the present work exhibited some advantages on quick procedures, less sample required, and reliable results, which could provide a new method for the identification of traditional Chinese medicine with the imitations and adulterants, and the control of quality and origin.
Drugs, Chinese Herbal ; analysis ; chemistry ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Reproducibility of Results ; Spectrophotometry, Infrared ; methods ; Spectroscopy, Fourier Transform Infrared ; methods

OBJECTIVETo investigate the clinical value of intraoperative ultrasonography in cervical laminoplasty.

METHODSThirty patients underwent cervical laminoplasty for cervical compressive myelopathy in our hospital from April 2010 to April 2012. Spinal cord compression ratio was calculated in preoperative MRI axial images for every patient. The spinal cord decompression status was evaluated by intraoperative ultrasonography during cervical laminoplasty. The pre- and post-operative Japanese Orthopaedic Association Scoring System (JOA) score was recorded, and the recovery ratios of surgery were graded using the Hirabayashi equation. The relationship between the parameters and neural recovery was explored.

RESULTSThe intraoperative ultrasonography was used to evaluate the ventral compression of the spinal cord. The ultrasonographic dynamic viewings were classified into three types based on the spinal cord contact with ventral structures after decompression: Type 1, non-contact (n=2); Type 2, contact and apart (n=16); and Type 3, contact (n=12). The patients were divided into two groups: group A, showing Type 1 or 2 findings, representing satisfied decompression; and group B, showing Type 3 findings with insufficient decompression. The recovery ratio was (73.3±21.0)%(31.3%-100.0%) in group A, but decreased to (43.2±33.1)%(0-83.3%) in group B(t=3.05,p=0.005). Pearson analysis showed that the patient age(r=-0.294,p=0.122), preoperative JOA score(r=0.059,p=0.759), and spinal compression ratio(r=0.269,p=0.151) was not correlated with Hirabayashi recovery ratio.

CONCLUSIONIntraoperative ultrasonography during laminoplasty is a feasible and promising method for evaluating spinal cord decompression status and predicting the prognosis following surgery.

Adult ; Aged ; Aged, 80 and over ; Decompression, Surgical ; Female ; Humans ; Laminectomy ; methods ; Male ; Middle Aged ; Monitoring, Intraoperative ; methods ; Spinal Cord Compression ; diagnostic imaging ; surgery ; Ultrasonography

Objective To determine the changes of interleukin-12(IL-12) and IL-12 mRNA in gastric mucosa of children with helicobacer pylori (Hp) infection,and to study the effects of Hp infection on the expression levels of IL-12 and IL-12 mRNA,and to evaluate its possible roles in the pathogenesis of gastric mucosal inflammation in Hp related gastroduodenal diseases.Methods Biopsy specimens were taken from the antral mucosa on endoscopy in patients with or without Hp infection, which were diagnosed by urease test and Giemsa staining. The expression levels of IL-12 and IL-12 mRNA in gastric mucosa were determined by ELISA and RT-PCR.Results Inflammation of gastric antral mucosa was more severe in Hp-positive mucosa .The expression levels of IL-12 and IL-12 mRNA in Hp-positive mucosa were (66.42?15.15) ng/g and (59.21?15.03)%,which were more than those in (Hp-negative )(22.22?8.79) ng/g and (17.94?7.39)%(P

Objective To examine the relationship between CCT 5 ,γδ T cell and autoimmune diseases .Methods Recombinant CCT5 protein was cloned , expressed and purified in E.coli.Three peptides of CCT5 protein were used to prepare for anti-CCT5 monoclonal antibodies .Purified CCT5 protein was used to expand γδT cells from pe-ripheral blood mononuclear cells (PBMC).Plasma level of CCT5 in healthy donors, patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) were detected by ELISA assays .The correlation analysis between plasma CCT5 concentration and the percentage of different subtypes of γδT cells measured by flow cytometry was made . Results The CCT5 gene was amplified by PCR and the length of the target fragment was 1 750 bp.The expressed 65 ku CCT5 protein was purified and validated by SDS-PAGE.Two paired monoclonal anti-CCT5 antibodies were screened to detect CCT5 protein in plasma.Immobilized recombinant CCT5 protein was able to induce specific sig-nificant amplification of peripheral γδT cells.Correlation analysis of 10 healthy donors indicated significant corre-lation between the plasma CCT 5 concentration and the proportion of Vγ9 and Vδ2 γδ T cells.The plasma CCT5 concentration significantly decreased in autoimmune diseases patients , including RA and SLE .Conclusions These data suggest that CCT 5 could be a novel Vγ9δ2 γδT cell-related factor in autoimmune diseases , which deepen the understanding of Vγ9δ2 γδT cell function in autoimmune diseases .

BACKGROUND:Pedicle screw is the major instrumentation of surgery in thoracic spine. However, there have been few reports about pedicle morphology relevant to screw insertion tracts, and few reports comparing the normal adolescents and adolescent idiopathic scoliosis patients. OBJECTIVE:To compare the morphologic characteristics of the thoracic pedicle with regard to safe thoracic pedicle screw placement in normal adolescents and adolescent idiopathic scoliosis patients. METHODS: Thoracic pedicles of thirty-five normal adolescents and thirty-five adolescent idiopathic scoliosis patients were measured with three-dimensional reconstruction CT images. Measured parameters include (1) critical distance: the shortest distance from an entry point to the ventral cortex of the lamina. (2) Safe distance: the distance from the entry point to the tangent of the spinal canal at the medial wal of the pedicle. (3) Pedicle screw length. (4) Pedicle width. (5) Pedicle transverse angle. The dangerous area was defined as the distance between the critical distance and the safe distance. RESULTS AND CONCLUSION: The mean critical distance was (9.2±1.0) mm for the normal adolescents, and (9.4±1.2) mm for the adolescent idiopathic scoliosis patients. Safe distances were significantly less in normal adolescents (14.7±0.8) mm than that of the adolescent idiopathic scoliosis group (15.4±1.4) mm (P < 0.001). The dangerous area was (5.4±0.7) mm for the normal adolescents, which was significantly less than that of the adolescent idiopathic scoliosis patients (6.0±1.0) mm (P < 0.001). Pedicle screw length was (36.6±4.1) mm for the normal adolescents and (37.1±5.3) mm for the adolescent idiopathic scoliosis patients. Pedicle width was (5.8±1.2) mm for the normal adolescents and (5.7±1.7) mm for the adolescent idiopathic scoliosis patients. No significant difference in critical distance, pedicle screw length and pedicle width was found between the two groups (P=0.382, 0.135, 0.293). Pedicle transverse angle decreased gradualy from T1 to T12 in both groups. These results verify that pedicle morphology of many parameters is different between normal adolescents and adolescent idiopathic scoliosis patients, especialy in the apical area of the thoracic curve.