BACKGROUND: The attack of temporal epilepsy is associated with the loss and death of hippocampal neurons, in which the specific pattern and mechanism of the loss of hippocampal neurons are still unclear, and it is hard to make sure the inevitable association of the epileptic discharge with activation of cysteine-containing ASPartate-specific protease (caspase 3)and neuronal apoptosis, of hippocampal neurons.OBJECTIVE: To observe the neuronal apoptosis and caspase 3 gene expression of in vitro cultured rat hippocampal neurons of epilepsy models.DESIGN: An open experiment.SETTINGS: Department of Neurosurgery, Changhai Hospital, the Second Military Medical University of Chinese PLA; Department of Neurosurgery,Changzheng Hospital, the University.MATERIALS: The experiments were carried out in the Neurosurgery Laboratory of the Second Military Medical University of Chinese PLA from June 2002 to June 2003. Ten male or female SD rats with 24 hours after birth were used. The Caspase 3 flow detection kit was purchased from American BD Company, and polymerase chain reaction (PCR) primers were synthetized by Shanghai Haojia Company.METHODS: ① The SD rats within 24 hours after birth were killed by cutting down the head to remove the brain, then bilateral hippocampi were taken out, and hippocanpal neuron models of epileptic discharge were established. The discharge of the models was recorded with whole cell patch clamp technique. The neurons cultured for 8 days and treated with Mg-free medium were taken as epileptic discharge model group, and those cultured for 8 days but not treated with Mg-free medium were taken as the blank control group, and the changes of potentials were recorded. ② The fulllength cDNA of caspase 3 was cloned with reverse transcription-polymerase chain reaction (RT-PCR), and then it was labeled. The expression of caspase 3 gene and neuronal apoptosis were detected with in situ hybridization and flow cytometry.MAIN OUTCOME MEASURES: ① Results of cDNA cloning of caspase 3; ② Results of Caspase 3 in situ hybridization; ③ Results of apoptosis.RESULTS: ① The products amplified by RT-PCR showed DNA segment lanes of about 800 bp after treated with 12 g/L agarose gel electrophoresis (Figure 1), which was concordant with the predicted value. The detection of DNA sequence showed that the length of the obtained cloning open-reading frame was 843 bp. ② The hybridization showed that in the blank control group, the positively stained hippocampal neurons were less than 10%, the neurites were well-stacked, and formed extensive synaptic association; In the epileptic discharge model group, the positively stained neurons were obviously increased at 3 hours after the Mg-free treatment, and there were many strongly and positively stained neurons at 12 hours, all these neurons kept the neurites, which became little. ③ The flow cytometry showed that at 6 hours after the Mg-free treatment, the apoptotic cells began to increase obviously, the numbers of apoptotic cells in certain times were not the same.CONCLUSION: Epileptic discharge can trigger the caspase 3 gene expression, by which neuronal apoptosis is induced.

Objective To clone the full-length of human Gill cDNA and construct the recombinant lentiviral expressing vector pLOX-Gfil for eukaryotic expression,providing a basis for further study on the biological functions of Gfil.Methods Total RNA was isolated from K562 cells,and the full-length Gfil cDNA was amplified by RT-PCR and then ligated with pGEM-T vector after retrieve and purification.The ligation product was transformed into competent cells DH5a.The positive recombinant clones were selected and identified by a complementation,restriction endonuclease digestion.The cloning vector and the lentiviral vector pLOX first digested with BarnH I were ligated and transformed.The enzyme and PCR analyses were performed to confirm the recombinant vector,and then DNA sequence analysis.Results A fragment of 1.2 kb was obtained by RT-PCR.The enzyme and PCR analyses revealed that the correct Gfil cDNA was cloned.The sequence of cloned cDNA was identical to the sequence deposited in GenBank (NM005263).Conclusion Gfil was cloned correctly and the recombinant lentiviral vector pLOX-Gfil for eukaryotic expression was constructed successfully.

Objective To explore the clinical curative effect and complications of digital three-dimensional molding of titanium mesh for repairing skull defect. Methods The clinical data of 42 patients having underwent repairing skull defect with three-dimensional molding of titanium mesh were retrospectively analyzed. Results The average operation time was about 2 h, and 42 patients were operated successfully. The bilateral skull was symmetry and the appearance was good. In 42 patients, subcutaneous dropsy occurred in 1 case, epilepsy occurred in 3 cases, and intracranial bleeding again surgery occurred in 1 case. Patients were satisfied with the results of cranioplasty. Conclusions Repairing skull with digital three-dimensional molding of titanium mesh is simple, with shorter operation time, lower operation risk, and lower postoperative complication, and the clinical curative effect is satisfactory.

Objective Off-pump coronary artery bypass (OPCAB),minimally invasive direct coronary artery bypass (MIDCAB) and robotic-assisted coronary artery bypass (RA-CAB) are all used to treat isolated left anterior descending artery (LAD) disease.The aim of this study is to compare the early outcomes after these three procedures.Methods From February 2009 to May 2012,102 consecutive patients underwent revascularization of LAD.31 patients were treated by OPCAB,45 by MIDCAB and 26 by RA-CAB.Patients received sternotomy in the OPCAB procedures.The MIDCAB procedures were performed through a 10-cm anterolateral muscle-sparing minithoracotomy.In the RA-CAB procedures,left internal mammary arteries (LIMA) were harvested with the aid of da Vinci surgical system and sewing of the anastomoses was performed under direct vision by a 3-cm anterolateral minithoracotomy.Results No significant difference was observed in graft flow,pulse index,renal failure,reoperation for hleeding,new onset of arterial fibrillation and deep wound infection between these three groups.There was also no significant difference in peri-operative mortality,major adverse cerebro-cardiovascular events (MACCE) between these three groups.Compared with OPCAB,MIDCAB and RA-CAB significantly reduced the need of blood transfusion (4.4％ vs.32.3％,P＜ 0.05; 7.7％ vs.32.3％,P＜0.05).The patients receiving RA-CAB had shorter length of postoperative stay than whom receiving OPCAB[(8.8 ± 3.2) days vs.(12.4 ± 7.7) days,P ＜ 0.05)].There is no significant difference between the outcomes of MIDCAB and RA-CAB.Conclusion These findings indicated that MIDCAB and RA-CABwere feasible,effective and safe options for revascularization of isolated LAD disease.MIDCAB and RA-CAB showed the advantage of less invasive and faster recovery,compared with OPCAB.Therefore,MIDCAB and RA-CAB should be the routine treatment for patients with isolated LAD disease.In some advanced centers,RA-CAB will be the preferred method.The mid-and long-term outcomes of these three methods should be further investigated.

In many pathogens infection,especially virus,antibody-dependent enhancement(ADE) can aggravate the infection and lead to severe diseases.In this immunopathological phenomenon,virus-specific antibodies enhance the entry of virus into monocytes,macrophages and granulocytic cells and even the replication of virus through different mechanism.This phenomenon has been reported in numerous pathogens including virus,bacteria and parasite and the mechanisms of ADE vary from different species.Further study of ADE can promote the vaccine research and development to make the most use of vaccine and prevent human body from pathogens,which will be helpful to control the spread of pathogens including Zika virus.In the present review,we review the research progress of ADE mechanism in recent years,including antibodies mediating,receptors mediating,complement mediating,viral proteins mediating and cellular mediating ADE.In addition,dengue virus,human immunodeficiency virus,Coxsackie virus,Ebola virus,Zika virus and other pathogens will be illustrated respectively.This review provides insights on the different mechanism of ADE in different pathogens.

Prostate cancer is a complex, heterogeneous disease that mainly affects the older male population with a high-mortality rate. The mechanisms underlying prostate cancer progression are still incompletely understood. Beta-adrenergic signaling has been shown to regulate multiple cellular processes as a mediator of chronic stress. Recently, beta-adrenergic signaling has been reported to affect the development of aggressive prostate cancer by regulating neuroendocrine differentiation, angiogenesis, and metastasis. Here, we briefly summarize and discuss recent advances in these areas and their implications in prostate cancer therapeutics. We aim to provide a better understanding of the contribution of beta-adrenergic signaling to the progression of aggressive prostate cancer.

This study investigated the association between serum 25(OH)D 3 levels and cardiovascular risk-related indicators. 4 727 participants aged 20 and above from the National Health and Nutrition Examination Survey 2015-2018 database were enrolled. Body mass index, hypersensitive C-reactive protein, high density lipoprotein cholesterol, systolic blood pressure, waist-height ratio, and total cholesterol were selected as the research indicators. Weighted multiple linear regression models, subgroup analyses, smooth curve fitting, and saturation threshold effect analyses were employed to explore the relationship between serum 25(OH)D 3 and these indicators. The results showed that after full adjustment for covariates, every 1 nmol/L increase in serum 25(OH)D 3, the changes in β (95% CI) values for body mass index(BMI), hypersensitive C-reactive protein(hs-CRP), systolic blood pressure(SBP), waist-height ratio(WHtR), high density lipoprotein cholesterol(HDL-C), and total cholesterol(TC) were -0.05 (-0.06, -0.04) kg/m 2, -0.01 (-0.02, -0.01) mg/L, -0.02 (-0.04, -0.01) mmHg, -0.000 7 (-0.000 8, -0.000 6), 0.10 (0.08, 0.11) mg/dl, and 0.08 (0.04, 0.12) mg/dl, respectively. Female participants were more sensitive to changes in serum 25(OH)D 3, while participants aged 60 and above were relatively less sensitive. The relationship between serum 25(OH)D 3 and these indicators partially exhibited nonlinear patterns across different gender and age subgroups. The saturation threshold effect analysis revealed 8 meaningful inflection points. In summary, vitamin D has a close association with cardiovascular risk-related indicators.

This study investigated the association between serum 25(OH)D 3 levels and cardiovascular risk-related indicators. 4 727 participants aged 20 and above from the National Health and Nutrition Examination Survey 2015-2018 database were enrolled. Body mass index, hypersensitive C-reactive protein, high density lipoprotein cholesterol, systolic blood pressure, waist-height ratio, and total cholesterol were selected as the research indicators. Weighted multiple linear regression models, subgroup analyses, smooth curve fitting, and saturation threshold effect analyses were employed to explore the relationship between serum 25(OH)D 3 and these indicators. The results showed that after full adjustment for covariates, every 1 nmol/L increase in serum 25(OH)D 3, the changes in β (95% CI) values for body mass index(BMI), hypersensitive C-reactive protein(hs-CRP), systolic blood pressure(SBP), waist-height ratio(WHtR), high density lipoprotein cholesterol(HDL-C), and total cholesterol(TC) were -0.05 (-0.06, -0.04) kg/m 2, -0.01 (-0.02, -0.01) mg/L, -0.02 (-0.04, -0.01) mmHg, -0.000 7 (-0.000 8, -0.000 6), 0.10 (0.08, 0.11) mg/dl, and 0.08 (0.04, 0.12) mg/dl, respectively. Female participants were more sensitive to changes in serum 25(OH)D 3, while participants aged 60 and above were relatively less sensitive. The relationship between serum 25(OH)D 3 and these indicators partially exhibited nonlinear patterns across different gender and age subgroups. The saturation threshold effect analysis revealed 8 meaningful inflection points. In summary, vitamin D has a close association with cardiovascular risk-related indicators.

Yersinia pestis, the cause of plague, is transmitted by flea bite. Y. pestis forms a biofilm in the proventriculus of its flea vector to enhance transmission. Biofilm formation in Y. pestis is positively regulated by the intracellular levels of the second messenger cyclic diguanylate (c-di-GMP). The c-di-GMP in Y. pestis is synthesized by two diguanylate cyclases (DGC), HmsT and HmsD, and degraded by phosphodiesterase (PDE), HmsP. Here we summarized the regulators that modulate c-di-GMP metabolism and biofilm formation in Y. pestis and discussed their regulatory mechanism.

OBJECTIVETo screen for KIT gene mutations in two Han Chinese pedigrees affected with Piebaldism.

METHODSClinical data of the pedigrees was collected. Genomic DNA was extracted from blood samples collected from the pedigrees and 120 unrelated healthy controls. All coding exons of the KIT gene were subjected to PCR amplification and direct sequencing.

RESULTSTwo missense mutations, c.1861G>A(p.Ala621Thr) and c.1872G>A(p.Met624Ile), were identified respectively in the two pedigrees. Neither mutation was found among healthy members from the respective pedigree and the 120 unrelated healthy controls. c.1872G>A is a novel mutation.

CONCLUSIONMutations of the KIT gene may affect the structure and function of the transmembrane receptor KIT, which lead to the disease.