ABSTRACT:Objective To study the BDNF/TrkB signaling pathway in the epilepsy model of hippocampal neurons and the regulatory effect of on it.Methods The primary hippocampal neurons cultured in vitro for 7 days were randomly divided into seven groups:control group,epilepsy group,control+BDNF group,epilepsy+BDNF group,control + miR-185 ? group,epilepsy + miR-185 ? group,and epilepsy + miR-185 ? + BDNF group.We constructed miR-185 ? lentivirus vector and observed the changes of BDNF/TrkB pathway expression after transfaction of miR-185 ? by immunohistochemistry,patch clamp technique and Western blot technique.Results Compared with the control+BDNF group,the phosphorylated TrkB (pTrkB)/TrkB value was significantly lower in epilepsy+BDNF group (P < 0.05 )and control group (P < 0.001 ).Compared with the epilepsy group,the phosphorylated TrkB (pTrkB)/TrkB value was significantly higher in epilepsy+BDNF group (P <0.05).Compared with the epilepsy+miR-185 ? +BDNF group,the phosphorylated TrkB (pTrkB)/TrkB value was significantly lower in epilepsy + BDNF group and epilepsy + miR-185 ? group (P < 0.001 ).BDNF could promote the signaling conduction and miR-185 ? could remove the inhibition of BDNF/TrkB signaling.Conclusion BDNF can activate the BDNF/TrkB signaling pathway and transfection with miR-185 ? can relieve the inhibition of BDNF/TrkB signaling pathway of epileptic state by up-regulating the expression of TrkB.

Objective:To investigate the correlation between polymorphisms of serine hydroxymethyltransferase1 gene and the adverse reactions of high-dose methotrexate (HD-MTX) in children with acute lymphoblastic leukemia (ALL). Methods:A total of 51 patients with ALL were treated with HD-MTX, and clinical manifestations after HD-MTX treatment were evaluated retrospectively. cD-NA was obtained from mRNA. The polymorphisms of SHMT1 gene containing rs1979277, rs3783, rs1979276, and rs12952556 sites were tested by denaturing gradient gel electrophoresis and direct sequencing. Effects of SHMT1 gene polymorphisms on HD-MTX ad-verse reactions were evaluated. Results:Severe adverse reactions in ALL patients treated with HD-MTX appeared to be mainly neutro-penia and hepatoadverse reactions. The frequency distributions of rs3783 (C>G), rs1979276 (C>T), rs12952556 (A>G), and rs1979277 (C>T) were the same. The polymorphisms of rs1979277 showed no correlation with neutropenia (P>0.05) but rs1979277 CT and TT genotypes were correlated with hepatoadverse reactions (CT: OR=0.129, 95% CI: 0.020 to 0.817, P=0.03; TT: OR=0.103, 95% CI:0.017 to 0.620, P=0.013). Conclusion: No correlation was found between the combination of rs1979277, rs3783, rs1979276, rs12952556, and neutropenia, but one or more of these loci may reduce the risk of hepatoadverse reactions.

Objective:To investigate the association between glutathione S-transferase pi (GSTP1) gene polymorphism and toxici-ties related to high-dose methotrexate (HD-MTX) in children with acute lymphoblastic leukemia (ALL). Methods:GSTP1 genotypes and allelic frequencies in 51 children with ALL were determined by Nest PCR, denaturing gel gradient electrophoresis (DGGE), and DNA sequencing. HD-MTX adverse reactions were analyzed using the National Cancer Institute Common Toxicity Criteria (NCICTC). Results:We identified three SNPs of GSTP1, including rs1695 (A313G), rs1138272 (G439T), and rs4891 (T555C). The wild types, het-erozygous types, and homozygous types of GSTP1 rs1695/rs4891 polymorphisms were detected in 32 cases (62.7%), 16 cases (31.4%), and 3 cases (5.9%), respectively. GSTP1 rs1695/rs4891 polymorphisms included only one heterozygous type and one homozygous type. The allele frequencies of the three SNPs were 21.6%, 2.9%, and 21.6%. The AG+GG/TC+CC genotype of GSTP1 rs1695/rs4891 was associated with decrease in the odds of peripheral hemoglobin (OR=0.25, 95%CI=0.06-1.00, P=0.049). The AG+GG/TC+CC genotype of GSTP1 rs1695/rs4891 in standard and intermediate-risk ALL children was significantly correlated with higher odds of gastrointesti-nal toxicity (OR=0.125, 95%CI=0.02-0.78, P=0.026). Conclusion:GSTP1 rs1695 (A313G)/rs4891 (T555C) gene polymorphism is as-sociated with the reduction of peripheral hemoglobin in ALL children and with the odds of gastrointestinal toxicity in standard and inter-mediate-risk ALL children who receive high-dose methotrexate.

Abstract： Objective　To retrospectively analyze the investigation and disposal of the COVID-19 outbreak caused by the transmission of the Omicron variant in infected imported cases, and provide basis for COVID-19 outbreak management. Methods　The description epidemiological method was used to describe the COVID-19 outbreak in Sanya City from March 31 to April 15, 2022. The propagation chain was mapped and the experience gained and shortcomings identified in emergency responses were analyzed. Results　The outbreak resulted in 95 reported locally transmitted COVID-19 cases with a incubation period M(P25, P75) of 4 (3, 5) d. In the 95 cases, the proportion of cases detected through close contact screening, centralized isolation, community screening, control area screening, active treatment (examination), and key population screening were 33.68%, 22.11%, 18.95%, 12.63%, 6.32%, 4.21% and 2.11%, respectively. The epidemic spread for 6 generations, causing 5 clusters of outbreaks and 12 cases of cluster disease. The epidemic affected 12 villages/neighborhood committees, 1 bar, 1 hospital, 1 small clinic, 1 farmer's market, 1 large shopping mall and 1 restaurant in 2 districts of Sanya City. The result of gene sequencing was Omicron variant BA.1.1. Through the immediate launch of emergency plans, nucleic acid and antigen testing, controlling close contact between infected persons and close contacts, suspending indoor business sites, central urban control, and temporary suspension, COVID-19 was controlled within 16 days. Conclusions　The transmission chain of this outbreak was clear and was caused by imported cases. Strengthening the management of the pass, doing a good job in information sharing and docking, timely screening for cases, screening, pushing, controlling high-risk groups, and implementing comprehensive control measures, can effectively prevent the spread of the epidemic, providing a reference for the control of epidemic situations in relevant scenarios.

Objective To assess the influence between managements in emergency room(ER) andoutcome of severe traumatic brain injury (TBI),in order to provide inference for treatment.Methods A retrospective analysis was performed in severe TBI patients and recorded next indexes.(1) The managements in ER,including endotracheal intubation and oxygenation,fluid resuscitation,and mannitol intake.(2) The vital signs arriving at ICU,including systolic pressure and blood oxygen saturation.(3) Prognostic indicators including inhospital mortality and days during ICU,the scores of Glasgow outcome scale (GOS) at discharge and 6 months after injury.Results In 140 severe TBI patients,65 patients (46.4％) died during ICU.The mortality of patients with endotracheal intubation [65.0％ (39/60)] was significantly higher than that without endotracheal intubation [32.5％(26/80)](P＜ 0.01).The mortality in whether fluid resuscitation and using mannitol had no significant difference [44.7％ (46/103) vs.51.4％ (19/37),49.2％ (31/63) vs.44.2％ (34/77)] (P ＞0.05).In days during ICU,there was no significant difference among the three treatment measures (P＞ 0.05).In GOS grade at discharge and 6 months after injury,the proportion of 4 and 5 grade were 8.3％ (5/60) and 25.0％ (15/60) in patients with endotracheal intubation,while 27.5％ (22/80) and 52.5％ (42/80) in patients without endotraeheal intubation (P ＜ 0.01).In fluid resuscitation and using mannitol patients,there were no significant difference(P ＞ 0.05).Conclusion Treating severe TBI patients in ER,endotracheal intubation should be carefully chosen,fluid resuscitation and mannitol may not be given.

OBJECTIVETo study the correlation of tenascin-c (TN-C) degradation with relapse and/or metastasis in stage-I non-small cell lung cancer (NSCLC) in order to search for a potential biomarker for predicting recurrence, and also to investigate the molecular mechanism of TN-C degradation. Methods The fragment of TN-C in 63 surgically treated stage-I NSCLC was detected by Western blotting, and the activity of matrix metalloproteinases (MMPs) was also examined by gelatin zymography.

RESULTSTN-C degradation fragment was positively detected in 12 of 63 patients, and 9 of these 12 patients (75.0%) were found to develope recurrence during follow-up. The recurrence-free survival at 4 years was 28.1% in patients with positive TN-C degradation versus 82.1% in those without (P < 0.001), and which was 76.6% at 10 years in the patients without TN-C degradation. The activity of MMP-2 in the patients with positive TN-C degradation was also found to be significantly higher than that in the patients without (P < 0.001).

CONCLUSIONTenascin-c degradation fragment may be a reliable biomarker for predicting recurrence and/or metastasis in the early NSCLC, and matrix metalloproteinase-2 may be a responsible proteinase for degradation of tenascin-c.

Aged ; Aged, 80 and over ; Biomarkers, Tumor ; Bone Neoplasms ; metabolism ; secondary ; Brain Neoplasms ; metabolism ; secondary ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; metabolism ; pathology ; surgery ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Pneumonectomy ; Tenascin ; metabolism

BACKGROUNDGap junction channels formed by connexin43 (Cx43) protein are important in cardiac morphogenesis, and Cx43 gene is thought to be associated with congenital heart malformation (CHM). This study was undertaken to detect the mutations of Cx43 in fetuses with CHM.

METHODSCx43 extron DNA was amplified by PCR from 16 fetuses with a variety of CHM. The PCR products were analyzed by SSCP and DNA sequencing. Thirty children who had no CHM were selected as controls.

RESULTSEight homozygous mutations of Cx43 were observed in a fetus with double outlet right ventricule (DORV), five of the 8 mutations were missense mutations including Arg239Trp, Ser251Thr, Ala253Pro, Pro283Leu and Thr290Asn, and the remaining 3 were silent polymorphisms including Gly252Gly, Pro256Pro and Thr275Thr. No mutations were found in other fetuses and the control group.

CONCLUSIONSMutations of Cx43 may be associated with congenital conotruncal anomalies. PCR-SSCP is an effective method for screening the mutations of Cx43.

Connexin 43 ; genetics ; Fetus ; metabolism ; Heart Defects, Congenital ; genetics ; Humans ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Sequence Analysis, DNA

Receptors play a crucial role in determining the host specificity and tissue tropism of virus. Foot-and-mouth disease virus(FMDV)has been showed to use four integrins, ?v?1, ?v?3, ?v?6 and ?v?8 as receptors to initiate infection and ?v?6 functions as the major receptor.The cDNA encoding bactrian camel integrin ?6 from the lung tissue was cloned and sequenced. The 2367bp cDNA of bactrian camel integrin ?6 encodes a polypeptide of 788 amino acids consisting of a 26-residue putative signal peptide, a 681-residue ectodomain with 8 potential N-linked glycosylation sites and 58 cysteine residues, a 29-residue transmembrane domain, and a 52-residue cytoplasmic domain with a NPLY motif and 1 potential N-linked glycosylation site. The nucleotide sequence similarity of integrin ?6 between bactrian camel and cattle, pig, sheep, human, mouse, Norway rat is 91.1%、91.8%、90.6%、90.5%、83.7%、84.1%, and the amino acid sequence similarity is 94.3%、93.4%、93.4%、93.7%、88.7%、88.6%, respectively. The bactrian camel ?6 gene exhibited the higher sequence homology with the ?6 gene of cattle, pig and sheep, indicating their close genetic relationships. It is possible that host tropism of FMDV may related to divergence in ?6 receptors among different species.

This study was purposed to investigate the expression and clonal proliferation of receptor (TCR) Vbeta subfamilies of the T-cells in acute leukemic patients at different disease status (onset, complete remission or relapse) and to analyze the influence of the leukemic cell load on anti-leukemic effect of peripheral T-lymphocytes of the patients. Gene sequences of peripheral TCR Vbeta 24 families from 11 leukemic patients and 3 normal donors were expanded by RT-PCR. Genescan technique was applied to evaluate clonal expression of the TCRVbeta subfamilies, clonal characteristics of the CDR3 from peripheral blood of AML patients at different disease status. The application, clonal proliferation, cellular complexity of T-cells, and the variation of immunotypes of T-cells were compared. The results indicated that the lower and partial distribution of TCR Vbeta subfamily was found in all 11 patients when firstly diagnosed; the expression of TCR Vbeta subfamilies after induction in vitro increased; obvious elevation of TCR Vbeta subfamilies was observed in patients at complete remission although expression level was still lower than normal, whereas the significant descent of TCR Vbeta subfamilies was detected in 4 relapsed patients. Only 1 - 2 clonal proliferation of TCR Vbeta subfamilies existed in 9 out of 11 patients at initial diagnosis which increased at remission. The status of clonal proliferation of Vbeta subfamily T-cells continued regardless of any different disease status in most patients. There was an obvious decrease of CDR3 complexity at initial diagnosis or relapse, while CDR3 complexity would be partially improved at remission. It is concluded that the restrict distribution and expression of TCR Vbeta subfamilies were found in AML patients. Clonal proliferation of T-cells Vbeta subfamily continuously exists regardless of any different disease status in most patients. Some Vbeta subfamilies sustain clonal proliferation at different disease status. Some clonal proliferations of Vbeta subfamilies are associated with the effects of leukemic cells, CDR3 complexity obviously decreases under disease status which can be partially improved at remission.
Adult ; Aged ; Clone Cells ; Complementarity Determining Regions ; genetics ; Female ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Receptors, Antigen, T-Cell ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Young Adult

Receptors play a crucial role in determining the pathogenesis and tissue tropism of virus. Foot-and-mouth disease virus (FMDV) has been showed to use four integrins, alphavbeta1, alphavbeta3, alphavbeta6 and alphavbeta8 as receptors to initiate infection. In this study, the porcine integrin alphav gene was cloned by RT-PCR from the lung tissue of healed pig infected experimently with FMDV, and compared its nucleotide and deduced amino acid sequence with the av gene of other animals. The 3141bp cDNA of bovine integrin alphav encodes a polypeptide of 1046 amino acids consisting of a 30-residue putative signal peptide, a 955-residue ectodomain, a 29-residue transmembrane domain, and a 32-residue cytoplasmic domain. The ectodomain contains 11 potential N-linked glycosylation sites (NXT/NXS), 2 calcium binding domains (DX[D/N] XDGXXD) and 18 cysteine residues. The nucleotide sequence similarities of integrin alphav between pig and cattle, human, rheses monkey, house mouse, chicken, dog are 93.3%, 91.5%, 91.4%, 85.6%, 73.2% and 89.9% respectively; and the amino acid sequence similarities are 96.3%, 94.6%, 94.1%, 90.8%, 81.6% and 93.8%, respectively. The alphav gene of cattle and pig exhibited the highest sequence homology. It is possible that host tropism of FMDV may related to divergence in receptors among different species.
Amino Acid Sequence ; Animals ; Base Sequence ; Cattle ; Cloning, Molecular ; DNA, Complementary ; genetics ; Foot-and-Mouth Disease Virus ; physiology ; Integrin alphaV ; genetics ; Macaca mulatta ; Molecular Sequence Data ; Receptors, Virus ; genetics ; metabolism ; Sequence Analysis ; Swine ; genetics