Objective To discuss the value of Fisher discriminant analysis of serum progesterone and the growing rate of β-human chorionic gonadotropin in the prediction of early ectopic pregnancy. Methods 66 patients with ectopic pregnancy (11 cases were successfully treated expectantly and 55 cases were treated surgically including 40 cases of rupture of fallopian tube and 15 cases of tubal abortion) and 55 patients with intrauterine pregnancy and 50 patients with threatened abortion were chosen. Serum progesterone,β-HCG,48 hβ-HCG and the 48 h growing rate of β-HCG in each group were measured and a Fisher discriminant analysis was used. Results The serum progester-one was (30.27± 18.20) nmol/L in ectopic pregnancy group,( 108.44±23.27 ) nmol/L in intrauterine pregnancy group and (91.68±34.90) nmol/L in threatened abortion group. The first β-HCG was ( 3767.63 ± 3530.38 ) U/L in ectopie pregnancy group,(29 028.65 ± 10 874.01 )U/L in intrauterine pregnancy group and (13 457.47±16 367.65)U/L in threatened abortion group. The second β-HCG was (4349.24±3536.22)U/L in ectopic pregnancygroup,(56 139.46 ± 23 296.87 ) U/L in intrauterine pregnancy group and (23 270.63 ± 23 811.68 ) U/L in threat-ened abortion group. The growing rate of β-HCG ( β-HCG/the first serum β-HCG) was 1.29 ± 0.28 in ectopic preg-nancy group,1.93 ± 0.36 in intrauterine pregnancy group and 1.97±0.28 in threatened abortion group. There was significant difference in serum progesterone,the first β-HCG and the second β-HCG as well as the growing rate of β-HCG among the groups(P<0.05 or <0.01). Fisher discriminant analysis of combing progesterone and the growing rate of β-HCG were connected with diagnosis of ectopic pregnancy,however,the only one serum β-HCG was not con-nected with diagnosis of ectopic pregnancy. 98.5% of ectopic pregnancy,65.6% of intrauterine pregnancy and 64.0% of threatened abortion were correctly classified in the Fisher discfiminant analysis,with overall correct rate of 77.8%. Conclusion Fisher discriminant analysis of combing progesterone and the growing rate of β-HCG can bet-ter predict the early ectopic pregnancy.

Objective To investigate the effect of aldehyde-free fixation solution and aldehyde fixation solution on DNA and cytoplasm of nucleus.Methods The DNA quantification (IOD),discrete coefficient (CV),DNA index (DI) and nucleus area (area) were measured by DNA quantification system (ICM) in 8 different fixation solution;combined with cytological staining,the effects of different fixative solution on nuclear DNA and cytoplasmic staining were analyzed.Results Aldehyde fixation solution was better than aldehyde free fixation solution,the formaldehyde in fixtion solution has important influence on Feulgen-eosin staining.Conclusion Aldehyde fixation solution combined with cytological staining can obtained good dyeing effect,which provides the basis for the subsequent multi-technique joint diagnosis.

Objective: To illuminate the gene characteristics and clinical characterization of Coxsackievirus B5 (CV-B5) strains isolated from patients with sevre hand, foot and mouth disease (HFMD) in Qingdao city. Methods: A total of 1 844 patients of HFMD were consecutively admitted to Qingdao Women and Children's Hospital from 2013 to 2014. Information of the study population described above was collected retrospectively. The samples were collected from at least 1 site (throat swab, cerebrospinal fluid), which viral nucleic acid extracted and the entire VP1 gene sequences of CV-B5 isolates were amplified and sequenced, then the homology and phylogeny analysis were conducted by MEGA7.0. The prototype Faulkner strain and other VP1 amino acid sequences were derived from the GenBank database. Results: A total of 8 CV-B5 positive cases were obtained, including 4 males and 4 females; 6 severe hospitalized cases and 2 outpatients. The age of 6 hospitalized patients ranged from 3 to 48 months, with a median of 26 months. For the six inpatients, fever, convulsions vomiting, diarrhea and rash were the main clinical manifestation, and all combined with viral encephalitis. Compared with the prototype strain Faulkner, in the VP1 region,the nucleotide and the amino acid homologies was 77.3%-78.8% and 95.5%-97.0% respectively. Five out of the six severe cases with substitution of serine (S) to asparagine (N) at amino acid site 95 in the VP1 region. The sequences of 8 CV-B5 strains were classified into genogroup D. Conclusion Hand, foot and mouth disease associated with CV-B5 virus infection can result in nervous system involvement and the main complication was viral encephalitis. The CV-B5 strains associated with severe hand, foot and mouth disease had high nucleotide homology and present a certain regional aggregation.

Objective:To investigate the molecular features of hand, foot and mouth disease (HFMD) caused by coxsackievirus A10 (CVA10) in Qingdao and analyze the clinical features of mild and severe cases.Methods:A total of 6 677 cases of HFMD routinely monitored by Qingdao Women and Children Hospital from 2014 to 2021 were enrolled. Throat swab samples were collected. Clinical data of these cases were retrospectively analyzed. Virus nucleic acid was extracted from the samples and the serotypes of enteroviruses were identified. The VP1 genes of CVA10 strains were amplified and sequenced. A phylogenetic tree based on the VP1 gene sequences was constructed using MEGA7. 0 software. SPSS23.0 software was used for statistical analysis.Results:There were 285 cases positive for CVA10, including 183 males and 102 females, and children under five years old accounted for 89.8%. Most of CVA10 infection occurred between the months of April to September. The count of white blood cells, the percentage of neutrophils, the concentration of hemoglobin, and the levels of aspartate aminotransferase and alanine transaminase were significantly higher in severe patients than in mild patients. Besides, chest radiography and brain CT revealed more abnormalities in severe patients, and the duration of ECG monitoring was longer in them. Compared with mild cases, severe cases developed rash early than fever with rash mostly on buttocks ( P<0.05). Phylogenetic analysis showed that most of the CVA10 strains circulating in Qingdao between 2014 and 2021 belonged to clade Ⅰ, and there were two variations A23V and I283V in the amino acid sequence of clade Ⅰ. Conclusions:This study showed that children of all ages were susceptible to CVA10, especially those under five years old. CVA10 showed complex and diverse epidemic trends in different regions and years.

Objective:To investigate the etiological characteristics of hand, foot and mouth disease (HFMD) and the VP1 gene evolution of coxsackievirus A6 (CVA6) in Qingdao from 2020 to 2022.Methods:Total enterovirus (EV), enterovirus 71 (EV71), coxsackievirus A16 (CVA16), CVA6 and coxsackievirus A10 (CVA10) were detected by fluorescent RT-PCR in throat swab samples from HFMD patients in Qingdao from 2020 to 2022. The full-length sequence amplification of VP1 gene and gene sequence determination of CVA6 isolates were performed, and the phylogenetic and molecular characteristics were analyzed by MEGA-X software.Results:The overall positive rate of enteroviruses in 1 645 samples collected during 2020-2021 was 80.73%(1 328/1 645). The proportions of EV71, CVA16, CVA6, CVA10 and other enteroviruses were 0.08%(1/1 328), 19.65%(261/1 328), 68.15%(905/1 328), 0.98%(13/1 328) and 11.14%(148/1 328), respectively. Children under 5 years old accounted for 75.68% (1 245/1 645) of all cases and 100% (2/2) of severe cases. A phylogenetic tree was constructed based on the VP1 region sequences of 45 CVA6 strains showing all strains belonging to the D3a subtype. There were 10 non-synonymous mutations of amino acids in the VP1 protein by gene evolution analysis.Conclusions:CVA6 D3a subtype is the main epidemic strain causing HFMD in Qingdao from 2020 to 2022, especially in children under 5 years old.