Humans ; Polyomavirus ; classification ; Polyomavirus Infections ; virology

Objective To explore the effect of cryopreserved canine kidney cells (MDCK)single -layer on the isolation and proliferation of influenza virus B.Methods Revived P17 MDCK cells were passage for 2 -4 gener-ations,and subsequently preserved in 4℃ refrigerator for 3,6 and 9 days,respectively.Under same experimental con-ditions,the 4℃ refigerater preserved cells were co -incubated with influenza -like illness(ILI)throat swab speci-mens.Cytopathic effect (CPE)was observed,and the proliferation of virus was determined using real -time PCR and the hemagglutinin titers were determined by serological test.Results (1)CPE:The CPE of the MDCK cells pre-served in 4℃ refrigerator for 3 or 6 days had no significant differences compared with that in the control group,while the cell preserved in 4℃ refrigerator for 9 days showed CPE fastly and maintained for a short time.(2)Real -time PCR:the proliferation of influenza virus B in the MDCK cells preserved with 4℃refrigerator for 3 or 6 days (25.86 × 105 -30.25 ×106 ,26.31 ×105 -30.54 ×106 )had on difference compared with that of the control group (24.82 × 105 -29.86 ×106 ),with the proliferation rate of 105 to 106 times,while the proliferation cell with 4℃ cryopreserved for 9 days(19.72 ×104 -28.34 ×105 )the proliferation in cells preserved for 9 days was sharply decreased,with pro-liferation rate of 104 to 105 times.(3)The HA titer:The virus strains with hemagglutination titer above or equal to 116 (P >0.05)isolated with MDCK cells preserved in 4℃ refrigerator 3 or 6 days were not significantly different from that of the control group (10.92 ±0.79).And the cells with 4℃ cryopreserved for 9 days were significantly dicreased (P <0.01).Conclusion No significant effects on the isolation and proliferation of influenza virus B using MDCK cell preserved in 4℃ frigerator for near one week were observed in the present study.

Objective:To understand the genetic variation and the prevalence of H9N2 subtype avian influenza virus in Guizhou province, and to provide the scientific evidence for the prevention and control of avian influenza virus.Methods:The results of AIV detection in live poultry market(LPM) environment in Guizhou province from October 2018 to March 2019 were statistically analyzed, RNAs were extracted and sequenced from the HA genes of 13 samples of H9N2 positive screened by real-time PCR. Then the homology, the genetic evolution and the mutations of important amino acid were analyzed by bioinformation softwares. Results:The positive rate of AIV was 52.2% and the positive rate of H9N2 was 83.7% in LPM environment. The homology between nucleotides of the HA gene of 21 strains ranged from 91.6% to 100.0%, and the homology between amino acids of the HA gene ranged from 91.0% to 100.0%. All strains belonged to Y280 sublineage and G57 genotype. Key sites analysis showed that they had a common motif PSRSSRGLF and LSRSSRGLF at the cleavage site, which indicated that they were lentogenic and low pathogenic strains. Mutations H191N, E198T/A and Q234L at the receptor binding sites in the HA was found in 21 strains, while indicated the viruses had the potential to bind human-like receptor. The analysis results of glycosylation motifs showed that all 21 strains had 7 glycosylation sites, but had a site deletion at amino acid site 218 and an addition at 313.There was no significant mutation in the key site compared with the human infected strains. Conclusions:The detection rate of AIV in LPM environment in Guizhou province was high, and the pollution was very serious, and H9N2 subtype is the main subtype, All H9N2 subtype AIVs belonged to Y280 sublineage and G57 genotype, and they were low pathogenic avian influenza viruses in Guizhou province, but the genetic gap were widening and mutations of key amino acid site might enhance susceptibility and pathogenicity to human beings. Hence, It is necessary to strengthen the surveillance of molecular characteristic variation of H9N2 subtype AIV.

Objective To investigate the association of cysteine aspartic acid specific protease 8 (CASP 8) gene-652 6N Insertion/Deletion polymorphisms and susceptibility of type 2 diabetes mellitus (T2DM). Methods CASP 8 gene -652 6N I/D polymorphisms were detected by polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing in 414 controls and 410 patients with T2DM. Results I/I, I/D and D/D genotype frequency were 56.5%, 38.9%, 4.6%in controls and 58.0%, 32.9%, 9.0%in T2DM group respectively (P<0.05). The risk in D/D genotype people was 1.916 times than I/I genotype (adjusted OR=1.916, 95%CI=1.199~3.054, P<0.05). The fasting blood sugar of D/D genotype people was significantly higher than that of I/D and I/I genotype people (P<0.05). Conclusions CASP 8 gene-652 6N I/D polymorphisms are associated with T2DM outbreak.

Objective To understand the genetic variations of influenza B virus outbreaks in Guizhou province in 2016,and to compare the matching situation of outbreak epidemic strains with the vaccine strains recommended by WHO and representative strains in China.Methods The haemagglutinin HA1 gene of 8 strains isolated from two episodes of influenza B virus outbreaks in Tongren area was amplified and sequenced.The sequencing products were analyzed by bioinformatics software DNAStar. Results The two episodes of influenza outbreaks were both caused by influenza B Victoria lineage virus (BV).The homologies of the isolated strains were 99.8%—100.0% in nucleotide and 99.5 %—100.0%in amino acid.Mutation was only detected at 274 site in some strains.Compared with reference strain B/Victoria/2/87,the homologies were 91 .8%—92.0% and 91 .5 %—92.0%,respectively.Mutations developed at 17 amino acid sites,among which,I143V,V163I and V201I site were associated with the main antigenic determinant area B,C and D.Compared with previous vaccine strain B/Brisbane/60/2008, the homologies were 98.2%—98.3% and 98.5 %—99.0%,respectively,and mutations were detected at 3 sites.Mutations at I143V and N155D were detected in all 8 strains and at T247I in some strains.The mutation of I143V was associated with antigenic determinant area B.Compared with the representative strain B/Chongqing-Yuzhong/1384/2010,the homologies were 96.7%—96.8% and 97.0%—97.5 %, respectively.A total of 6 sites developed mutations,among which,5 sites were P84L,I143V,N155D, V172I and T223N mutations.The mutation of T247I was detected in some strains,and I143V was associated with area B.Compared with the epidemic strain in Guizhou in 2016,the homologies were 99.8%—100.0% and 99.5 %—100.0%,respectively.Mutation was only detected at site 247 in some strains and was not associated with the main antigenic determinant area.Conclusions The two episodes of influenza outbreaks in Guizhou are caused by the same BV lineage epidemic virus strain.Haemagglutinin gene of BV lineage virus is constantly changing.However,there is no new mutation emerged at important site.Compared with previous influenza vaccine strain B/Brisbane/60/2008 recommended by WHO,BV lineage virus is well matched and could provide a positive protection effect.

Objective To understand the epidemic characteristics and regularity of influenza B virus in Guizhou Province, and provide scientific evidence for the control and prevention of influenza.Methods Results of reverse transcription polymerase chain reaction(PT-PCR) of influenza B virus in Guizhou Province from April 1, 2013 to March 31, 2016 were statistically analyzed.Results A total of 1 904 samples were detected influenza B virus by RT-PCR, B/Yamagata (By) lineage and B/Victoria (Bv) lineage were 1 215 and 642 respectively.In April 2013-March 2014 and April 2014-March 2015, the predominant strains of influenza B were both By lineage, in April 2015-March 2016, the predominant strains of influenza B were Bv and By lineages, the epidemic peaks were in winter and spring;there's a higher positive percentage of influenza B in male, accounting for 56.83%;the highest detection rate of influenza B virus was found in population aged <15 years(70.80%),Bv and By lineages were the highest in the 0~ (42.37%) and 5~ age groups (35.56%) respectively;the main pathogen causing mixed infection was By+Bv (67.65%),mixed infection with influenza B virus accounted for 95.59%.Conclusion There are two lineages By and Bv epidemic in Guizhou Province, the epidemic peaks of influenza B are in winter and spring, male cases are higher than female, people under 15 years old are the high-risk group for influenza B, it is of great significance to strengthen the vaccination and surveillance of influenza in low age population.

Objective To explore the therapeutic effects and dose of Topiramate(TPM)in children with Tourette syndrome(TS).Me-thods Seventy-nine children with TS were given oral TPM,which were treated with Topiramate [0.5-3.0 mg/(kg?d),twice a day].The therapeutic effects were assessed using Yale Global Tic Severity Scale(YGTSS)before and 3 months after treatment and the side-effects of the drugs were observed.Results The differences of YGTSS scores before and after treatment,motertic score(19.63?3.09 vs 5.05?1.74),vocaltic score[(18.95?2.56)vs(4.82?1.94)],global scole score[(24.21?5.89)vs(10.42?3.69)],severity score[(62.21?5.81)vs(22.26?4.81)],there were significant differences of every score of YGTSS between before and after treatment(Pa

Objective To investigate the molecular characteristics of H5 subtype avian influenza viruses (AIV) in Weining, Guizhou Province. Methods Nine representative strains were randomly select-ed from H5 subtype AIV that were identified by real-time PCR in Weining, Guizhou Province from 2015 to 2017. Nucleic acid was extracted from each sample and hemagglutinin (HA) genes were amplified and then sequenced. Homology, genetic evolution and the sites related to pathogenicity, receptor binding regions as well as potential glycosylation of H5 AIV were analyzed by bioinformation software. Results Homology analysis revealed that there was 96. 1%-99. 9% and 95. 7%-100% similarity among the nine strains in nu-cleotide and amino acid of HA gene, respectively. These strains belonged to two branches, H5-1 and H5-2. The cleavage site motifs were PLREKRRKR↓GLF for five strains in H5-1 branch and PQRERRRKR↓GLF for four strains in H5-2 branch, which made them high pathogenic. QSG and QRG at the key receptor bind-ing sites were found in H5-1 and H5-2 branch strains, respectively. They were responsible for receptor bind-ing specificity of AIV. Mutations of 138Q, 139G and 53K were all detected in the nine strains. 129K, 189T, 140K and 282V mutations were discovered in the five strains of H5-1 branch, while 189N, 140M and 282I mutations were found in the four strains of H5-2 branch. Results of the glycosylation motif analysis showed that six sites were conservative, but there was an addition of 124NHT site in two strains of H5-2 branch isolated in 2017. Conclusion Two high pathogenic H5 subtypes of AIV could be epidemic in Wein-ing, Guizhou Province during 2015 to 2017. Although H5 subtype AIV did not possess specific receptor binding regions like human influenza viruses, they were in continuous variation with an increase in glycosyla-tion motifs, which might enhance their virulence and pathogenicity to human beings. Hence, surveillance and study on the molecular properties of H5 subtype AIV should be strengthened.

The study was aimed to investigate the genetic background and proliferation characteristics of multiple myeloma (MM). Myeloma cells were isolated from bone marrow of 19 MM patients by direct immunomagnetic cell sorting and the DNA content and cell cycle analysis were carried out by flow cytometry. The results showed that in 4 patients the myeloma cells were found to be hyperdiploid and in 15 patients those were found to be diploid respectively by DNA content analysis; the proportion of plasm cells from normal controls in S + G(2)/M phase was (1.15 +/- 0.60)%, and that of myeloma cells from MM patients was (10.06 +/- 12.60)% which was significantly higher than that in the former (p = 0.001). The incidence of hyperdiploid in newly diagnosed patients was 11.76%, and that of treated patients was 100.00% which was significantly higher than that in the former (p = 0.035); the proportion of myeloma cells from newly diagnosed patients in S + G(2)/M phase was (7.12 +/- 4.98)%, and that of treated patients was (35.10 +/- 32.56)% which was also significantly higher than that in the former (p = 0.001). It is concluded that the variety of myeloma cells in DNA content and cell cycle suggests the complicated genetic background and abnormal proliferation of MM, which relate with the course of disease to some extent.
Adult ; Aged ; Aged, 80 and over ; Bone Marrow Cells ; metabolism ; pathology ; Cell Cycle ; genetics ; Cell Proliferation ; DNA ; analysis ; genetics ; Diploidy ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; pathology

OBJECTIVETo investigate the clinical significance of the deletion of the long arm of chromosome 13 [del (13 q) ] and translocation of immunoglobulin heavy chain gene [t (14 q) I in multiple myeloma (MM) patients.

METHODSMyeloma cells were isolated from hone marrow by direct immunomagnetic cell sorting and interphase fluorescence in situ hybridization (FISH) was performed in 24 MM patients to detect del (l3q) and t (l4q).

RESULTSThe positive rates of del (l3q) and t (l4q) were 45.83% and 37.50% respectively. Five patients (20.83%) had both two abnormalities and 15 patients (62.50%) had at least one abnormality. Univariate analysis showed that the positive rates of del (l3q) were 35.71% and 66.67% in responders and non-responders (P = 0.214) and the positive rates of t (l4q) were 21.43% and 66. 67% in responders and non-responders (P = 0.077). Multivariate analysis showed that del (13q) (OR = 5.761, 95% CI 0.500-66.391, P = 0.160), t (14q) (OR = 6.576, 95% CI 0.580-74.614, P = 0.129), and corrected serum calcium level (OR = 8.080, 95% CI 0.738-88.427, P = 0.087) were relatively independent negative factors for response to therapy, with the corrected serum calcium level being the strongest reversely-correlated factor.

CONCLUSIONSInterphase FISH is a sensitive method to investigate the cytogenetics of MM. Del (13q), t (14q), and corrected serum calcium level can be used to predict treatment response and prognosis.

Adult ; Aged ; Aged, 80 and over ; Chromosome Deletion ; Chromosomes, Human, Pair 13 ; genetics ; Chromosomes, Human, Pair 14 ; genetics ; Female ; Humans ; Immunoglobulin Heavy Chains ; genetics ; In Situ Hybridization, Fluorescence ; Interphase ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; Translocation, Genetic