Objective To investigate the changes of lymphocyte subsets before and after the treatment with operation and chemotherapy in patients with breast cancer.Methods Using cytometer to measure lymphocyte subsets of CD3+,CD4+, CD8+,CD4+/CD8+ and NK of 25 breast cancer patients before and after treatment.The all 25 patients were treated with operation and chemotherapy after pathological diagnosis.The article compared the figures of T cell subsets before and after treatment.Results Compared with before operation and chemotherapy,the CD8+ statistically decreased after 1~3 months of treatment (t=2.57),and NK statistically increased after 3~6 (t=-2.23).After 6 months of treatment,the patients’ CD3+(t=-2.25),CD4+(t=-2.07)were statistically lower and NK (t=-3.56)was statistically higher.There was no significant difference for other items.Conclusion Lymphocyte subsets test against breast cancer patients can reflect immuni-ty level at a certain level during treatment.But how to explain the figures in different stage against treatment effectiveness and prognosis need further research.

Objective Compare the HPV infection characters between women in different geographic area of Ankang (south of Shaanxi) and Yanan (north of Shaanxi),to provide evidence based reference for cervical cancer control.Methods PCR amplification in vitro combined reverse dot-blot hybridization assay were used to analyze HPV genotypes of 7 912 cases from Ankang City Center Hospital,and 3 543 cases from Yanan Affiliated Hospital of Yanan Medical University from June 2012 to July 2016,and compared HPV infection rate,age and hypotype distribution of the cases.Results The HPV infection rate of Ankang area was 21.75％ (1 721/7 912),and Yanan area was 41.41％ (1 402/1 721).The percentage of HPV positive women with high-risk hypotype was 81.46％ (1 402 1 721) in Ankang and 86.16％ (1 264/1 467) in Yanan,with low-risk hypotype was 11.04％ (190/1 721) in Ankang and 7.23％ (106/1 467) in Yanan,with high and low risk hypotype was 7.50％ (129 147) in Ankang and 6.61％ (97/1 467) in Yanan.The women more than 60 year old were higher HPV prevalence age group.The HPV infection rate of this age group was 30.41％ (104/342) in Ankang,and 50.00 ％ (60/120) in Yanan.Yanan's HPV infection rate was obviously higher than Ankang.The popular HPV hypotypes in Yanan were same with that in Ankang.The most popular high-risk hypotypes were HPV16,58 and 52,the most popular low-risk hypotypes were 6,11 and 81.Conclusion The HPV infection rate were rather high in all age group.The most popular HPV hypotypes were same between Ankang and Yanan area.But Yanan's HPV infection rate was obviously higher than Ankang (P＜ 0.01).The causes need further explore.

Objective To investigate the sequences of the 3' end of genomes from enterovirus 71 isolated from pediatric patients with different symptoms in Beijing during 2008-2009.Methods Clinical specimens were collected from pediatric patients suffering from hand,foot and mouth disease(HFMD)and/or with neurological complications who visited the affiliated Children's Hospital during the epidemic seasons of 2008 and 2009 in Beijing.The samples were inoculated into the Vero cell line,and the virus isolates were further identified by nested reverse transcription-nested PCR(RT-nPCR)assay using universal enterovirus primers,type specific primers for EV71 and CA16.The 3' end of genomes(including 3D and 3' UTR regions)from 10 EV71 strains derived from various clinical presentations were amplified and sequenced.Results Analysis of the nucleotide sequences of the amplified fragments showed that the 3' end of genomes of 10 EV71 isolates include the 3D region of 1386 nucleotides(nt)encoding the 3D polymerase(3Dpol,462 amino acids),the terminator codon TGA and 3' UTR of 81 nt.Nucleotide sequence identities among 3D regions from these 10 EV71 isolates were in the range of 95.8%-99.6%,while the nucleotide sequence identities for 3' UTR were 96.3%-100%.The majority of nucleotide changes were located at the third codon positions which caused silent mutations,thus the amino acid sequence changes of 3Dpol among those 10 EV71isolates were scanty.The residues 140 and 263 which were R and 1 were substituted by K and V,respectively in 3 of 4 neurovirulent strains,whereas only 1 of 6 strains from mild cases had these 2 amino acid changes.The sequences of the 3D and 3' UTR regions of 10 EV71 isolates were compared to the representative strains of known genotypes from the GenBank.The nucleotide and amino acid sequences of 10 EV71 isolates in the 3D region exhibited highest homology to the subgenotype C4 of EV71(92.7%-94.2%and 96.8%-97.6%.respectively).However,3' UTR of 10 EV71 isolates shared the highest nucleotide identity with CA16/G10(88.9%-91.4%).The phylogenetic analysis based on the 3D regions demonstrated that 10 EV71 isolates had the closest genetic relationship with the representative isolate of CA sub-genotype of EV71 and shared more homology with CA16/G10 than other known genotypes of EV71.Conclusion Genetic analysis of the 3' end of genomes from 10 EV71 strains indicated that the 3' end of genome may play a role in the evolution of EV71.

Objective To develop a convenient reverse transcription PCR(RT-PCR)method for identifying genotypes of human metapneumovirus(hMPV)from clinical samples.Methods According to the gene sequences of hMPV G with different genotypes,the A and B genotype specific primers were designed.A diplex RT-PCR was applied to identify different genotypes according to the molecular weight of PCR products in agarose gel.37 clinical samples were detected through this method.Results It was convenient to distinguish different genotypes of hMPV(383 bp for A and 284 bp for B)by the diplex RTPCR,and there was no non-specific amplification for common respiratory viruses.so it meant that the specificity of primers was good.The results of genotyping 37 clinical samples showed that 20 samples were identified as genotype A by both sequence analysis of M gene and diplex RT-PCR,whereas 17 samples were identified as genotype B by sequence analysis of M gene.but in these 17 samples 14 samples were identified as genotype B by the diplex RT-PCR and remaining 3 samples could not be genotyped because there was no PCR product after amplification.The consistency rate for these two methods Was 91.9%[(20+14)/37].Conclusion The method of diplex RT-PCR Was developed successfully and can be used for identify genotypes of hMPV.

Objective To investigate the mechanisms of action of adiponectin on receptor activator of NF-Kb ligand(Rankl) and osteoprotegerin (OPG)expressions in human osteoblasts.Methods Real-time PCR was used to detect the expressions of RANKL and OPG mRNA in cultured human osteoblasts. The phosphorylations of JNK, p38 mitogen-activated protein kinase (MAPK) , ERK1/2 were assayed by Western blot. RNA interference for adiponectin receptor, MAPK inhibitors SB203580 and SP600125 were used for elucidating the mechanism of the action of adiponectin in regulating OPG and RANKL expressions. Results Suppression of adiponectin receptor-1 (AdR1) expression with siRNA abolished the adiponectin-regulated expressions of OPG and RANKL mRNA in human osteoblasts. Furthermore, pretreatment of osteoblasts with MAPK inhibitor SB203580 abolished the expressions of adiponectin-regulated RANKL and OPG mRNA, but SP600125 did not show the effect. Conclusion Adiponectin induces the expression of RANKL and inhibits the expression of OPG in human osteoblasts through AdR1/p38 MAPK pathways.

Objective: To investigate the serotype and drug resistance of 815 Salmonella isolates from Hunan Province, so as to provide insights into management of Salmonella infections. Methods: Salmonella isolates were collected from stool samples of foodborne diarrheal patients and food samples in Hunan Province from 2020 to 2021, and serotyped. Antimicrobial susceptibility test was performed using the broth microdilution method. Results: A total of 10 groups and 39 serotypes were characterized in 815 Salmonella isolates. Among the 646 Salmonella isolates of human sources, 388 isolates were identified as serogroup B (60.06%), with S. typhimurium and its variants aspredominant serotypes (364 isolates, 56.35%), and among 169 foodborne isolates, 61 isolates were characterized as serogroup B (36.09%) with S. london as the predominant serotype (26 isolates, 15.38%). There were 597 antimicrobial resistant Salmonella isolates of human sources, with a drug resistance rate of 92.41%, and the percentage of ampicillin resistance was 81.58%. There were 140 foodborne antimicrobial resistant isolates, with a drug resistance rate of 82.84%, and the proportion of tetracycline resistance was 72.78%. However, Salmonella isolates from both humans and foods were sensitive to imipenem. In addition, there were 577 multidrug resistant Salmonella isolates, including 490 multidrug resistant isolates of human sources and 87 foodborne multidrug resistant isolates. Conclusions S. typhimurium and its variants and S. london were predominant serotypes of Salmonella isolates from 815 foodborne diarrheal patients and food samples in Hunan Province from 2020 to 2021, and a high rate of multidrug resistance was detected.

Background Acute respiratory infection (ARI) is one of the most common infectious diseases in infants and young children globally.This study aimed to determine the virus profile in children with ARI presenting with different severities.Methods Clinical specimens collected from children with ARI in Beijing from September 2010 to March 2011 were investigated for 18 respiratory viruses using an xTAG Respiratory Viral Panel Fast (RVP Fast) assay.The Pearson chisquare analysis was used to identify statistical significance.Results Of 270 cases from three groups of ARI patients,including Out-patients,In-patients and patients in the intensive care unit (ICU),viruses were detected in 176 (65.2％) specimens with the RVP Fast assay.The viral detection rate from the Out-patients group (50.0％) was significantly lower than that from the In-patients (71.1％) and ICU-patients (74.4％) groups.The virus distribution was different between the Out-patients group and the other hospitalized groups,while the virus detection rate and distribution characteristics were similar between the In-patients and ICU-patients groups.The coinfection rates of the Out-patients group,the In-patients group,and the ICU-patients group were 15.6％,50.0％ and 35.8％,respectively.In addition to respiratory syncytial virus (RSV) and adenovirus (ADV),human rhinovirus (HRV) was frequently detected from children with serious illnesses,followed by human metapneumovirus (hMPV),human bocavirus (HBoV) and coronaviruses.Parainfluenza virus 3 (PIV3) was detected in children with lower respiratory illness,but rarely from those with serious illnesses in the ICU-patient group.Conclusion In addition to so-called common respiratory viruses,virus detection in children with ARI should include those thoucht to be uncommon respiratory viruses,especially when there are severe ARI-related clinical illnesses.

OBJECTIVETo analyze the genotype, epidemic pattern and the characteristics of the disease of enteroviruses during the epidemic season of hand, foot and mouth disease (HMFD) in children from 2013 to 2014 in Beijing to provide the scientific evidence for prevention and treatment of HFMD.

METHODDuring April to September in 2013 and March to October in 2014, a total of 977 throat swabs were collected from children who visited the Children's Hospital Affiliated to Capital Institute of Pediatrics, including 147 from patients with HFMD in 2013, 343 with HFMD, 201 with atypical HFMD, 83 with herpangina, 25 with fever with convulsions, 64 fever with rash and 114 with rash in 2014. Enteroviruses universal type (EV), Enteroviruses type 71 (EV71) and Coxsackievirus group A 16 (CA16) were detected by real-time RT-PCR respectively. The nucleic acid of specimens which were identified with non-EV71, non-CA16 was tested by nested PCR and analyzed by VP1 sequencing. The detection rate and epidemic pattern of different genotypes of enterovirus were analyzed among different age groups and between 2013 and 2014.

RESULTOf 977 throat swabs, 80. 1% samples were detected positive for enteroviruses. The positive rates of CA16, EV71, CA6, CA10, CA4 and other EVs were 25. 6% (250/977), 18. 9% (185/977), 20. 0% (195/977), 5. 0% (49/977), 1.5% (15/977) and 9.1% (89/977), respectively. Twenty six of the 89 other EVs included CA2, CA5, CA8, CA9, CA12, CA14, CB2, CB5, E6, E9 and E25, each genotype of which was no more than 3. The nucleotide homologies shared among CA6, CA10 and CA4 strains between 2013 and 2014 were 94. 3% - 100%, 93. 8% - 99. 1% and 92.7% - 99. 8%, respectively. The positive rates of ≤1 year group were 71. 1% (106/149), which was lower than that of other age groups (all P <0. 05), but similar to that of >5 year group (χ2 =1. 181,P = 0. 277). In 2013, the positive rate of EV was 85. 7% (126/147) and the predominant genotype was CA6 54. 8% (69/126), followed by CA16 20. 6% (26/126) and EV71 11. 9% (15/126). In 2014, the positive rate of EV was 85. 4% (293/343) in the 343 children with HFMD, the predominant genotypes were CA16 with the positive rate of 42. 7% (125/293), EV71 with 38. 2% (112/293) and CA6 with only 11. 3% (33/293). In 2014, the positive rates of EV in 201 atypical HFMD, 83 herpangina, 25 fever with convulsions, 64 fever with rash and 114 rash were 83. 6% (168/201), 80. 7% (67/83), 76. 0% (19/25), 64. 1% (41/64) and 60. 5% (69/114), respectively. All genotypes of enteroviruses peaked mainly during May to August every year, but there were no obvious epidemiological pattern about each genotype.

CONCLUSIONCA6 became the main causative agent of HFMD in 2013, however, CA16 and EV71 predominated again in 2014 in Beijing. The clinical manifestations caused by CA6, CA10, CA4 and other genotype of enteroviruses differed from EV71 and CA16. Besides EV71 and CA16, more attention should be paid to CA6, CA10, CA4 and other type of enteroviruses.

Beijing ; epidemiology ; Child, Preschool ; Enterovirus A, Human ; classification ; Enterovirus Infections ; epidemiology ; virology ; Exanthema ; Fever ; Genotype ; Hand, Foot and Mouth Disease ; epidemiology ; virology ; Humans ; Infant ; Real-Time Polymerase Chain Reaction

Objective To investigate the etiological agents of hand, foot and mouth disease (HFMD) in children in spring and summer from 2007 to 2008 in Beijing and the characteristics of the disease by virus isolation and to provide the scientific evidence for prevention and treatment for HFMD. Methods During April to August, 2007 and May to September, 2008, 356 clinical specimens including 255 throat swabs and 101 vesicle fluids were collected from 256 patients with HFMD who visited the Children's Hospital Affiliated to Capital Institute of Pediatrics and children with severe HFMD with neural system complications from Ditan Hospital and Youan Hospital All of the specimens were inoculated into Vero cells for virus isolation. After the cell pathogenic effects (CPE) appeared, the isolates were identified by RT-PCR with the universal primers within 5'untranslated region of enterovirus and typed by specific primers for VP1 gene of EV71 and CA16, respectively. The throat swabs from all of 10 severe HFMD were tested for enterovirus by RT-PCR addition to virus isolation. Results Out of 256 patients, 188 were positive for enterovirus by virus isolation, with the overall positive rate of 73.4%. Among the 356 clinical specimens collected from these 256 patients, 239 enterovirus strains were isolated with the overall positive rate of 67.1%. The positive rate for virus isolation from vesicle fluid samples was 75.2% which was higher than the positive rate of isolation from throat swabs (63.9%), but the time for CPE appearing in cell culture showed no significant difference. The positive rate of virus isolation from throat swabs from children with severe HFMD was 50% (5/10) which was lower than overall positive rate (73.4%) from regular HFMD. The RT-PCR typing for virus isolates revealed that among 45 enterevirus strains isolated from the specimens collected in 2007 by the universal primer pairs, 43 were CAI6 (95.6%, 43/45) and 2 were EV71 (4.4%, 2/45), whereas for the specimens collected in 2008, out of 143 enterovirus isolates by PCR with universal primers, 117 were EV71 (82.4%, 117/142) and 24 were CA16 (16.8%, 24/142). All of 10 severe cases were positive for EV71 by RT-PCR directly from clinical specimens. Conclusion CA16 and EVT1 were the etiological pathogens of HFMD in Beijing during 2007 to 2008 HFMD seasons. The dominant type of enterovirus was different between 2007 and 2008. Enterovirus type CA16 was predominant in 2007, whereas EV71 was predominant in 2008. All of severe cases of HFMD in children in this study were caused by EV71.

Objective To investigate the status and clinical and epidemiological characteristics of human metapneumovirus (hMPV) and human bocavirus (HBoV) infections in children with acute respiratory tract infections (ARTIs) in Taiyuan. Methods A total of 549 children with ARTIs from November 2012 to May 2013 and November 2013 to May 2014 were recruited. The pharyngeal swab specimens were collected. The hMPV and HBoV were detected by using real-time PCR. Results In 549 children, 56 children (10.2%) were hMPV positive on swab specimens, 15 children (2.7%) were HBoV positive on swab specimens. The detection rates of hMPV and HBoV in November 2012 to May 2013 were 12.3%and 2.0%, respectively, and in November 2013 to May 2014 were 6.5%and 4.0%, respectively. The detection rate of hMPV was signiifcantly different between two periods (P<0.05), while the detection rate of HBoV has no signiifcant difference between two periods. In different months, the detection rate of hMPV and HBoV showed no signiifcant difference. The highest detection rates of hMPV and HBoV were all in children younger than two years old. The highest detection rate of hMPV was in children with asthmatic bronchitis or bronchiolitis. Conclusion In Taiyuan, during the monitoring periods, the ARITS are associated with childhood hMPV and HBoV infection especially in infants and toddlers. hMPV is one of the most important pathogens in infants and toddlers with wheezing.