Objective:Human leukocyte antigen(HLA)B27 is a susceptibility allele of ankylosing spondylitis(AS),and HLA-B27 antigen typing is an important indicator for clinical diagnosis of AS,but current typing methods such as sequence specific primer polymerase chain reaction(PCR-SSP)still possess limitation.Therefore,this study aims to analyze the correlation between B27 subtypes and susceptibility to AS in Hunan Province by applying high-resolution polymerase chain reaction-sequence-based typing(PCR-SBT). Methods:Peripheral blood of 116 patients with suspected AS(suspected AS group)and 121 healthy volunteers(control group)admitted to the Second Xiangya Hospital from January 2020 to December 2020 were collected for HLA-B genotyping by PCR-SBT.Among the patients in the suspected AS group,23 patients were finally diagnosed with AS(confirmed AS group),and the remaining 93 undiagnosed patients served as the non-confirmed AS group.PCR-SBT and PCR-SSP were used to detect HLA-B27 typing in 116 patients with suspected AS,and the results of the 2 methods were compared. Results:The HLA-B27 allele frequency in the suspected AS group was significantly higher than that in the control group[11.63%vs 2.48%;P<0.001,odds ratio(OR)=5.18,95%confidence interval(CI)2.097 to 12.795].B*27:04,B*27:05,B*27:06,and B*27:07 were detected in the suspected AS group and the control group.The frequency of the B*27:04 allele in the suspected AS group was significantly higher than that in the control group(9.48%vs 1.24%;P<0.001,OR=8.346,95%CI 2.463 to 28.282).The positive rate of B27 in the suspected AS group and the confirmed AS group(B27+/+ and B27+/-)was significantly higher than that in the control group(χ2=16.579,P<0.001;χ2=94.582,P<0.001,respectively).Among the confirmed AS group,21 were HLA-B27 carriers,and the B27 positive rate in the confirmed AS group was 91.3%.PCR-SBT could achieve high resolution typing of the HLA-B gene locus,with higher sensitivity,specificity,positive predictive value,negative predictive value,and accuracy than PCR-SSP. Conclusion:PCR-SBT typing analysis shows a strong correlation between HLA-B * 27:04 and AS in Hunan province.The PCR-SBT method can be used as the preferred option for the auxiliary diagnosis of clinical AS.

Klebsiella pneumoniae(K.pneumoniae)is an important pathogen that can cause severe hospital-and community-acquired infections.To systematically investigate its methylation features,we determined the whole-genome sequences of 14 K.pneumoniae strains covering varying serotypes,multilocus sequence types,clonal groups,viscosity/virulence,and drug resistance.Their methy-lomes were further characterized using Pacific Biosciences single-molecule real-time and bisulfite technologies.We identified 15 methylation motifs[13 N6-methyladenine(6mA)and two 5-methylcytosine(5mC)motifs],among which eight were novel.Their corresponding DNA methyl-transferases were also validated.Additionally,we analyzed the genomic distribution of GATC and CCWGG methylation motifs shared by all strains,and identified differential distribution pat-terns of some hemi-/un-methylated GATC motifs,which tend to be located within intergenic regions(IGRs).Specifically,we characterized the in vivo methylation kinetics at single-base resolu-tion on a genome-wide scale by simulating the dynamic processes of replication-mediated passive demethylation and MTase-catalyzed re-methylation.The slow methylation of the GATC motifs in the replication origin(oriC)regions and IGRs implicates the epigenetic regulation of replication initiation and transcription.Our findings illustrate the first comprehensive dynamic methylome map of K.pneumoniae at single-base resolution,and provide a useful reference to better understand epigenetic regulation in this and other bacterial species.

[Abstract] Objective: To observe the short-term efficacy and safety of Apatinib combined with radiotherapy and concurrent docetaxel and cisplatin chemotherapy in driver-gene-negative non-small cell lung cancer (NSCLC) patients with brain metastases. Methods: A total of 72 NSCLC patients with brain metastases, who were treated in our hospital from June 2018 to June 2019, were enrolled in this study. The driver gene was proved to be negative by next generation sequencing (NGS). The patients were divided into control group (36 cases) and treatment group (36 cases) by Digital random grouping method.The control group received 2 cycles of chemotherapy with docetaxel and cisplatin and concurrent radiotherapy for brain metastases, and the treatment group was given Apatinib anti-angiogenic treatment based on the regimen in control group. Primary study endpoints: confirmed objective response rate (cORR) and disease control rate (DCR); Secondary study endpoints: progression-free survival (PFS), quality of life (QOL) score, serum carcinoembryonic antigen (CEA), vascular endothelial growth factor (VEGF), and incidence of adverse drug events (AE). Results: Compared with the control group, cORR and DCR in treatment group were significantly improved [41.67% (15/36) vs 33.33% (12/36), 80.56% (29/36) vs 69.44% (25/36), all P<0.05], the median PFS was significantly prolonged (5.9 vs 4.6 months, P<0.05), and serum CEA and VEGF levels were significantly reduced [(16.5±2.3) vs (22.9±3.7) ng/ml, (291.6±42.6) vs (479.3±50.2) ng/L, all P<0.05], while the QOL score was slightly increased, but the difference was not statistically significant [(69.5±8.5) points vs (64.1±7.3) points, P>0.05]. There was no statistically significant difference in the incidence of acute brain edema, gastrointestinal reaction, bone marrow suppression, and liver dysfunction between the two groups of patients (all P>0.05); however, the incidences of oral mucositis, hand-foot syndrome, hypertension and proteinuria in the treatment group were significantly higher than those in the control group (all P<0.05). Conclusion: The efficacy of Apatinib combined with radiochemotherapy in driver-negative NSCLC patients with brain metastases is significantly better than that of radiochemotherapy alone, and the adverse reactions can be controlled. It is worthy of clinical recommendation.

Objective To compare the effect of intensity modulated radiotherapy (IMRT) and conventional radiotherapy (CRT) combined with intracavitary brachytherapy on patients with cervical stump cancer. Methods A total of 47 patients with cervical stump cancer who were admitted to the First Hospital of China Medical University from October 2007 to September 2017 were retrospectively reviewed. Of the 47 patients, 19 patients received CRT combined with intracavitary brachytherapy (CRT group), and 28 patients received IMRT combined with intracavitary brachytherapy (IMRT group). In order to reduce the effects of selection bias and confounding factors, propensity score matching was performed to compare the groups. Results After the propensity score matching, there were 19 patients in each group. The 1-, 3-and 5-year overall survival (OS) rate for matched patients were 100.0％, 85.1％ and 63.8％ in IMRT group, and 84.2％, 57.9％ and 42.1％ in CRT group, respectively, and the difference in survival between the two groups was statistically significant (PLog-rank= 0.029, PCox= 0.043, HR= 3.723, 95％CI 1.044-13.280). The median progression-free survival (PFS) time of the IMRT group was better than that of the CRT group [median PFS time had not been reached (7-72 months) vs. 17 months (2-125 months), PLog-rank= 0.032, PCox= 0.042, HR=2.773, 95％CI 1.037-7.417]. The incidence of late rectal and bladder radiation injury was 15.8％ (3/19) and 0 (0/19) in IMRT group, and 57.9％ (11/19) and 26.3 ％(5/19) in CRT group, respectively, and the difference was statistically significant (P= 0.017, P= 0.046). Conclusion Compared with CRT combined with intracavitary brachytherapy, IMRT combined with intracavitary brachytherapy has a better effect on patients with cervical stump cancer, and the incidence of complications is low, this method is worthy of clinical application.

Cervical stump cancer is rare in clinic, which leads to insufficient understanding of it by clinicians. Radiotherapy alone is often used in the early stage, but the treatment mode has gradually changed recently. However, there is still no uniform treatment standard for residual cancer, and the prognostic differences of residual cancer patients under the new treatment mode have not been reported in real time. In order to improve the management of cervical stump, this article reviewed its clinical characteristics, treatment and prognosis.

Objective To investigate the pathogenesis of white matter damage (WMD) and the effects of xenon intervention on the expression of EphB4 and EphrinB2 mRNA in the brain tissue of neonatal rats.Method Three-day-old SD rat pups (n =96) were randomly assigned into sham group (n =24),model group (n =24),xenon intervention group 1 (n =24) and xenon intervention group 2 (n =24).The WMD model was established by injected of lipopolysaccharide (LPS) 0.05 mg/kg combined with ligation of the right carotid artery for 1 h in the last three groups.Rats in xenon intervention group 1 inhaled 50％ xenon immediately for 3 h after modeling,while rats in xenon intervention group 2 inhaled 50％ xenon for 3 h at 2 h after modeling.After the completion of xenon intervention,6 rat pups in each groups were sacrificed at 0 h,24 h,48 h and 72 h.The pathologic examination of periventricular tissue was conducted with hematoxylin-eosin staining (HE) and the expression of EphB4 and EphrinB2 mRNA was assayed by real-time quantitative polymerase chain reaction (RT-PCR).Statistical analysis was then performed.Result (1)The structure of white matter in model group became loose,band net-like,with significant nucleus pyknosis.The pathological damages in xenon intervention group 1 and 2 were lighter at 24 h,48 h and 72 h than model group,with less karyopycnosis.(2) Compared with the sham group,the expressions of EphB4 and EphrinB2 mRNA at 0 h,24 h,48 h and 72 h were significantly higher in the model group and xenon intervention group 1 and 2 (P ＜ 0.05),except for the EphB4 mRNA in xenon intervention group 1 at 72 h (P ＞ 0.05).The expressions of EphB4 and EphrinB2 mRNA at each time point in xenon intervention group 1 and 2 were decreased significantly than the model group (P ＜ 0.05),except for the EphB4 mRNA in xenon intervention group 2 at 72 h (P ＞ 0.05).However,there was no statistically significant difference on EphB4 and EphrinB2 mRNA between two xenon intervention groups at each time point (P ＞ 0.05).Conclusion The expression of EphB4 and EphrinB2 mRNA are appreciably increased in brain tissue of neonatal rats with WMD,which indicates the reactive angiogenesis.The intervention with xenon may play a neuroprotective role through reducing the expressions of EphB4/EphrinB2 mRNA and angiogenesis,and early intervention may be better.

Objective To analyze etiology,clinical manifestation and diagnosis process of vertigo and dizziness in children.Method The clinical data of 82 children with vertigo and dizziness treated in our hospital from January 2006 to December 2016 were retrospectively analyzed.Results There were 46 girls and 36 boys with a female:male ration of 1.28:1.The median age of patients was 9.0 years (14 months-18.0 years) and 21 cases were less than 6 years old.The chief complaints were repeatedly falling,crying,walking unsteadily and scratching the ear for unknown reasons.For children aged ＞ 5 years may express " roof or tent rotation";for those aged ＞ 6-＜ 15 years (n =34),the main complaint was more ambiguous "dizziness";for those aged ≥ 15-≤ 18 years (n =27) with the main complaint of " dizziness" may clearly express the " sense of rotation" or " the feeling of feeling drowsy,the top-heavy sense," and the accompanying deafness,earfullness,tinnitus and so on.Among 82 cases,there were 15 cases of benign paroxysmal vertigo (BPV,18.3％),12 cases of secretory otitis media (SOM,14.6％),11 cases of vestibular migraine (VM,13.4％),9 cases of benign paroxysmal positional vertigo (BPPV,11.0％),8 cases of inner ear malformation (9.8％),8 cases of Meniere's disease (9.8％),7 cases of vestibular neuritis (8.5％),6 cases of sudden deafness with vertigo (7.3％),4 cases of central vertigo (4.9％),1 case of rare genetic disease (1.2％) and 1 case of vertigo due to mental psychological (1.2％).The incidence of BPV,VM and Meniere's disease in girls was higher than that in boys.There was a tendency to self heal in BPV with the age increasing,in 15 BPV cases,8 after 12 years of age,3 after 14 years old had no vertigo attacks,and 4 cases were still in follow-up.The attack frequency of VM was decreased,and the extent was reduced with the age.Children older than 6 years were able to cooperate to vestibular function tests,and the majority completed the tests.Conclusion The analysis shows that the etiology of vertigo and dizziness in children is different from that of adults.Central vertigo and rare genetic disorders with dizziness as the first symptom are of great harmfulness,so intensive observation and multidisciplinary consultations are recommended.

Objective To analyze the genotypes and homology of MSP?1 and CSP gene of Plasmodium vivax in Shandong Province,so as to provide the evidence for case traceability. Methods A total of 12 blood samples were collected from P. vivax?infected cases in Shandong Province in 2011. Parasite genomic DNA was extracted. Primers were designed according to MSP?1 and CSP gene sequences of P. vivax. Then Nested PCR,enzyme digestion,sequencing and sequence alignment,and homolo?gous analysis were performed. Results The MSP?1 gene of all the 12 samples from P. vivax?infected cases were detected with a 470 bp PCR amplification band,and 350 bp and 120 bp enzyme digestion fragments,which were identified as type Sal?1. An analysis of phylogenetic tree of MSP?1 gene showed that the sequences of 9 indigenous case samples in Shandong Province were located in the same branch,one case sample infected from India was located in the same branch with India strains. All the 12 P. vivax?infected samples covered GDRA(D/A)GQPA sequences in CSP gene,which were identified as type PV?Ⅰ. Of the CSP gene among 12 P. vivax?infected samples,10 samples of indigenous case in Shandong Province and one sample of the case in?fected in Guangdong Province were detected with both 560-840 bp and 150-230 bp PCR amplification bands,which were iden?tified as temperate zone family strain of type PV?Ⅰ. However,one sample from the case infected in India was detected only with a 560-840 bp band,which was identified as tropical zone family strain of PV?Ⅰ. An analysis of phylogenetic tree of CSP gene showed that the sequences of 10 samples from the indigenous cases in Shandong Province and one sample from the case infected in Guangdong Province were located in the same branch,one sample from the case infected in India was located in the same branch with India and Indonesia strains. Conclusion Of all the indigenous isolates in Shandong Province,MSP?1 gene is geno?typed type Sal?1,CSP gene is genotyped temperate zone family strain of type PV?Ⅰ,with a high homology found among the in?digenous isolates.

Objective: To investigate the prognostic value of dynamic monitoring of RUNX1-RUNX1T1 transcript in pediatric patients with t (8;21) acute myeloid leukemia (AML) . Methods: The clinical features and RUNX1-RUNX1T1 transcript levels of 55 pediatric t (8;21) AML patients, newly diagnosed from Jan. 2010 to Apr. 2016, were analyzed retrospectively. The relationship between the minimal residual disease (MRD) and prognosis was analysed by dynamic monitoring of RUNX1-RUNX1T1 transcript levels using real-time quantitative PCR (RQ-PCR) technology. Results: The RUNX1-RUNX1T1 transcript levels in bone marrow cells at diagnosis was not related to relapse. After one course of induction therapy, patients with a more than 2 Log reduction of RUNX1-RUNX1T1 transcript levels (>2 Log) had lower 5 years cumulative incidence of relapse (CIR) [ (24.3±8.4) % vs (52.6±9.7) %, χ²=9.046, P=0.003], relapse-free survival (RFS) [ (71.6±12.7) % vs (48.1±13.2) %, χ²=5.814, P=0.016], and better overall survival (OS) [ (76.9±12.5) % vs (48.9±14.7) %, χ²=6.346, P=0.012], compared to patients with a less than 2 Log reduction (a<2 Log) . Multivariate Cox survival analysis suggested that a>2 Log reduction in RUNX1-RUNX1T1 transcript levels after a course of induction therapy was an independent prognostic factor for RFS (HR=0.263, 95%CI 0.081-0.851, P=0.026) and OS (HR=0.214, 95% CI 0.057-0.808, P=0.023) . During consolidation therapy and follow-up period, molecular relapse of 16 cases and hematologic relapse of 13 cases were identified by continuous dynamic monitoring of RUNX1-RUNX1T1 transcript levels, with a median interval of 4.0 (1.5-5.8) months from the molecular relapse to hematologic relapse. 2 cases of molecular relapse who received timely allogeneic hematopoietic stem cell transplantation did not experience hematologic relapse. Conclusion Dynamic monitoring RUNX1-RUNX1T1 transcript levels by RQ-PCR technique can subdivide patients into relatively low and high risk group, early screen patients at high risk of relapse and provide a scientific basis for precision stratification and risk-adapted therapy for pediatric t (8;21) AML children.

Objective To determine the expression of indole-2,3 dioxygenase (IDO) in human acute leukemia,and to investigate its correlations with clinicopathological parameters and prognosis in acute leukemia.Methods The expression of IDO in protein and RNA levels was detected by immunohistochemistry and real-time quantitative RT-PCR,respectively,and the correlations of IDO with clinicopathologic features and prognosis of acute myeloid leukemia (AML)-M5 were analyzed.Results The positive rate of IDO protein was 63.3 ％ (38/60) in human acute leukemia,while it in AML (34/49,69.4 ％),especially in AML-M5 patients (29/35,82.9 ％),was significantly higher than that of acute lymphoblastic leukemia (4/11,36.4 ％).The expression of IDO protein in healthy human peripheral blood mononuclear cells was negative.The RNA expression level of IDO in AML-M5 or non AML-M5 patients were significantly higher than that of healthy people (P ＜ 0.001),and AML-M5 patients had significantly higher IDO RNA level than that in non AML-M5 patients (P ＜ 0.05).The IDO gene expression was not correlated with sex,age and drug sensitivity,while it was closely related with these factors in the patients without complication of pulmonary infection.IDO could not act as an independent prognostic marker.Conclusion The expression of IDO in AML-M5 patients is significantly higher than that in non AML-M5 patients and healthy people.The positive expression of IDO is associated with poor prognosis of AML-M5 patients,but it is not an independent poor prognostic indicator.