Objective To investigate the influence of lentivirus mediated short hairpin RNA(shRNA)target to human papilloma virus(HPV)16 E6 on invasive ability of cervical cancer Caski cells.Methods Lentivirus was produced after shRNA target to human papilloma virus(HPV)16 E6 and to nonsense was cloned to lentivirus work vector.Infection ratio was assessed by assay of EGFP positive cells of Caski.Total mRNA of E6 was determined by RT-PCR after Caski cells were infected by lentivirus.The change of E6 protein expression was analyzed by Western blot.The invasive ability of Caski cells was assayed employing Transwell.Results The optimal MOI (Multiplicity of infection)of lentivirus to Caski was 2.5.Total mRNA and protein of E6 were decreased (by 70%and 63%)in interfering group compared with control group.The invasive ability of Caski cells also reduced after infected by lentivirus.Conclusion shRNA mediated by lentivirus can inhibit expression of HPV16 E6 and invasive ability of cervical cancer cells.

In order to explore the significance of enterovirus environmental surveillance of sewage,two sewage treatment plants in Fuzhou City were selected as sentinel surveillance sites.One sewage sample was collected from entrance of each plant every month,and total 48 samples were studied from 2013 to 2014 in Fuzhou City.Using the RD,L20B and Hep-2 cell lines to isolate the viruses,44 of the 48 sewage samples (91.67％) were positive,268 strains were isolated which covered at least 22 enterovirus serotypes.The peak season of virus isolation from sewage was in February and July which was two months earlier than patient surveillance.Eehovirus type 7 (Echo 7) was the predominant serotype in 2013 with 58 isolations (43.61％),and in 2014 echovirus type 6 (ECHO 6) became the predominant serotype with 85 isolates (62.96％),among the strains isolated from sewage sample.Phylogenetic analysis found that the predominant serotype Echo 6 and Echo 7 isolated from sewage were more diversified than that from clinical cases during the same period,and were high homology with isolates form patients which belong to same branch in phylogenetic tree.In conclusion,enterovirus environmental sewage surveillance is an effective approach,and could be more sensitive than patient surveillance.

OBJECTIVE To investigate the effect of overexpression of vascular cell adhesion molecule-1(VCAM-1)on the migration in vitro of the murine mesenchymal stem cells(MSCs)and its possible mechanism. METHODS The migration ability of normal mouse MSC (C3) ,empty vector-transfected MSC(C3+N) and VCAM-1 transfected MSC(C3+VCAM-1)was assessed by Transwell culture system in vitro after incubation for 8 and 12 h,respectively. The fetal bovine serum (FBS) was used as the chemotactic agent to induce MSC migration. The transmigrated cells were detected with methylosaniliam chloride(crystal violet)as well as DAPI staining.Furthermore,the specific chemical inhibitors of mitogen-activation protein kinase (MAPK) pathway ( SB203580,PD98059 and JNK inhibitorⅡ)were added to the Transwell system for 12 h and the alteration of the MSC migration ability was evaluated. RESULTS After incubation with FBS for 8 and 12 h,the absolute migrated cell number(7467 ± 485 and 8795 ± 255)and migration rate〔(14.9 ± 1.0)% and(17.6 ± 0.5)%〕of MSC in C3+VCAM-1 group were significantly increased compared with C3 group〔2731±562 and 4779±224, (5.5 ± 1.1)%and(9.6 ± 0.4)%〕and C3+N group〔2539 ± 321 and 5645 ± 1080,(5.1 ± 0.6)%and(11.3 ± 1.1)%〕(P<0.05,P<0.01),but there was no significant difference between C3 and C3+N groups. Moreover,the MSC migration ability of C3+VCAM-1 group was partially suppressed by addition of JNK inhibitorⅡ. The transmigrated cell number(4843 ± 167)and migration rate〔(9.7 ± 0.3)%〕were decreased compared with those of C3+VCAM-1 group without JNK inhibitorⅡ(P<0.01). SB203580 and PD98059,as specific chemical inhibitors of MAPK pathway,had no effect on MSC migration. CONCLUSION VCAM-1 can enhance mouse MSC migration in vitro and th4e mechanism may be related to JNK/MAPK pathway activation.

In this study ,street trains were isolated from brain of suspected rabid dogs via mouse inoculation technique (MIT) and cell culture inoculation technique (CIT) .Virus was identified by FAT ,RT-PCR and sequencing .Then ,the virus’ biological characteristics were analysed .The TCID50 test results demonstrated that viral titers were not high ,which showed the BHK-21 cells was not the best host cell of rabies viral strain .The LD50 test results showed it was a strong strain of rabies virus .These results laid a strong foundation for rabies laboratory research in Fujian .

The prevalence of acute flaccid paralysis (AFP) associated with EV71 and the genetic variation in Fujian , China from 2003 to 2012 was investigated in this study .Descriptive epidemiology was used to analyze the epidemiologic and clinical features of AFP cases associated with EV 71 .Phylogenetic analysis was performed to explore the genetical characteris-tics of EV71 based on the complete VP1 nucleotide and amino acid sequences .Results showed that the mean incidence of EV71-associated AFP in children under 15 years old was 2 .24/10 000 000 in Fujian Province during 2003 and 2012 ,based on the number of EV71 isolates and the reported AFP cases .And the incidence has increased since 2008 .The EV71 strains isolated from the AFP cases or from the healthy contacts were distributed in 9 prefectures of Fujian Province ,most in the months of May and June .Of 76 .0% (19/25) of AFP cases associated with EV 71 were the children under 3 years and the male-to-female ratio was 1 .5 :1 .Twenty out of twenty-two cases (90 .91% ) had fevers before the onset of paralysis .Most cases had unilater-al limb paralysis (14/22 ,63 .6% ) .Typical manifestations of hand-foot-and-mouth disease (HFMD) were observed in five cases before the onset of paralysis .Residual paralysis was observed in two cases during the follow-up visits .The strains isolated from 25 cases belonged to genotype C4 .All other strains belonged to subtype C4a except the subtype C4b strains isolated in 2003 .The homology among the strains was high in 2009-2011 ,and the homology among these strains and the representative strains in Fuyang ,Anhui Province was also in the high level .Therefore ,it was possible that the isolated strains had the same origin and might cause the epidemic .In conclusion ,an AFP surveillance system could be developed for analyzing the incidence of AFP associated with EV71 ,determining the features of the isolates ,and describing the intensity and trends of EV71 epidem-ics .

Objective To investigate the correlation between plasma soluble CD14 (sCD14)level and disease progression in patients with acute phase of acquired immunodeficiency syndrome (AIDS). Methods Forty-one human immunodeficiency virus (HIV)-infected patients were followed up from June 2007 to June 2010 in Beijing You′an Hospital,including 20 patients with CD4 + T lymphocyte counts more than 350/μL,and 21 less than 350/μL after 3 years of HIV infection.Twenty healthy blood donors were recruited as controls.Enzyme-linked immunosorbent assay (ELISA)was employed to test the plasma sCD14 level of healthy controls and patients infected with HIV at 1 -30 d,31 -90 d,91 - 180 d and 181 -360 d.Student t test was used to compare the means between two groups.ANOVA analysis was used to compare the means among more than two groups.Results The mean plasma sCD14 level in control group was (1 654±904)μg/L.Three years after HIV infection,the sCD14 level of patients with CD4 + T lymphocyte counts less than 350/μL group was (4 214±2 635)μg/L,which was higher than that of patients with CD4 + T lymphocyte counts more than 350/μL ([2 275 ±1 457 ]μg/L).The difference was statistically significant(t=-5 .41 ,P <0.01).The plasma sCD14 level in patients infected with HIV 181 -360 d was significantly higher than that in patients infected with HIV 1 - 30 days ([4 485 ± 2 779]μg/L vs [2 577 ±1 635 ]μg/L;t = -3.39,P <0.05 ).The plasma sCD14 level was positively correlated with HIV viral load (r =0.35,P =0.000 1 ),and negatively correlated with CD4 + T lymphocyte counts (r=-0.28,P =0.001 ).Conclusions The plasma sCD14 level in patients with acute phase of HIV infection is higher than that of healthy people,which increases with prolonged HIV infection.Plasma sCD14 level in of HIV infection acute phase may be closely related to HIV/AIDS progression.

Objective To explore the effect of percutaneous kyphoplasty (PKP) combined with Xianling Gubao capsule on treatment of osteoporotic vertebral compression fractures(OVCF).Methods 118 postmenopausal women diagnosed as OVCF and meeting the standard were collected from Mar.2012 to Jul.2016 in our hospital.They were randomly divided into two groups:observation group and the control group.General information of the two groups were comparable.The patients of the control group received PKP combined with traditional anti-osteoporosis therapy.The patients of observation group received Xianling Gubao capsule on the basis of the control group.The lumbar bone density (BMD,greater trochanter BMD,femoral neck BMD,Ward triangle BMD,the Japanese orthopaedic association(JOA)score,the visual analogue score (VAS) score,the serum levels of osteoprotegerin (OPG),receptor activator of nuclear factor-κB ligand (RANKL)and adverse reactions were recorded during the treatment.Results The lumbar BMD,greater trochanter BMD,femoral neck BMD,Ward triangle BMD of observation group at 6-month after operation were higher than those before operation and those of control group (P＜0.05).The VAS score and JOA score of the observation group were superior to those of the control group at 6-month after operation (P＜0.05).The serum levels of OPG of observation group were higher than those of control group,while the serum levels of RANKL of observation group were lower than those of control group at 6-month after operation (P＜0.05).No severe adverse reactions occurred in the two groups during the treatment.Conclusion Xianling Gubao capsule can significantly improve bone metabolism of postmenopausal women with osteoporotic vertebral compression fractures after percutaneous kyphoplasty,increasing BMD,improving lumbar function,relieving pain,without increasing complication rates.

Objective:To analyze the complete genome sequence characteristics of genotype G mumps virus isolated from Putian city, Fujian province in 2015.Methods:Two mumps virus strains isolated in Putian city, Fujian province in 2015 were selected for the study and named as MuV15-01 and MuV15-23.The whole genome of the isolated strains was amplified by reverse transcription-polymerase chain reaction (Rt-PCR). The genetic differences and antigen site variation of the isolated strain were analyzed by comparing the sequences of vaccine strain and other genotypes strains.Results:The total length of genome of MuV isolates was 15, 384 nucleotides. The nucleotide differences between isolates and other genotypes of mumps virus based on the whole genome ranged between 3.7% and 6.0%. The maximum and minimum difference was found between isolates and genotype A (the vaccine strains), and genotype B to N strains, respectively. Among the nucleotide sequences encoding different viral proteins, SH gene had the largest variation, while M and L genes were the most conserved. There were 7 and 11 amino acid mutations in the known antigen related sites of N gene and HN gene, respectively. Compared with the vaccine strain, the isolated strain added an additional N-glycosylation site to the 464 site of the HN gene.Conclusions:Genetic variations were detected in the epidemic mumps virus strains in Putian, Fujian, compared with other mumps genotypes and vaccine strains based on the whole genome. These resluts indicate that the monitoring for genetic differences between epidemic mumps strains and vaccine strains should be further strengthened.

Objective:To investigate the detection status and genotypes distribution characteristics of norovirus(NoV)in environmental sewage from three monitoring points in Fujian province, and to explore the significance of its application to NoV monitoring.Methods:Sewage samples were collected monthly at 5 sampling sites in representative monitoring cities, enriched and concentrated. Partial gene fragments of norovirus VP1 were amplified by reverse transcription-semi nested polymerase chain reaction (RT-snPCR), TA cloned and sequenced. Genotypes were identified based on the sequencing.Results:A total of 56 sewage samples were collected from July 2022 to June 2023. The detection rates of GⅠ and GⅡ were 89.29% (50/56) and 94.64% (53/56), respectively. A total of 7 NoV GⅠ genotypes and 13 GⅡgenotypes were identified. GⅠ.1, GⅠ.4, GⅡ.4 and GⅡ.17 were the dominant genotypes. NoV genotypes detected in different sampling sites were not exactly the same. The detection rate of NoV was low from August to November 2022, and the prevalence of the dominant genotypes was different in different seasons. GⅠ.1 and GⅡ.4 were highly prevalent from August to November 2022, but were replaced by GⅠ.4 and GⅡ.17 from December 2022 to June 2023, respectively. More NoV genotypes were detected in January-June 2023, comparing to the July-December 2022. The dominant genotype GII.17, has multiple clades and new variants have been discovered that are different from the 2014/2015 circulating strains.Conclusions:The detection rates of NoV in environmental sewage were very high, and genotypes were diverse. Environmental sewage surveillance could be an important complementary method for NoV cases surveillance.

Objective:To evaluate the effects of vaccine switch on the levels of neutralizing antibody (NA) of poliovirus in children aged<12 years.Methods:Subjects aged<12 years from 9 districts in Fujian province were chosen by stratified cluster and complete random sampling method. Blood samples were collected for testing NA of poliovirus by microcell neutralization.Results:A total of 2 134 subjects aged<12 years were selected. The positive rate of NA against PVⅠand Ⅲ were 98.64% and 95.83%; and geometric mean titers (GMTs) were 1∶259.35 and 1∶105.14, respectively. The GMTs presented a trend of decreasing as age increased. Compared to trivalent oral poliovirus vaccine (tOPV), bivalent oral poliovirus vaccine (bOPV) and inactivated poliovirus vaccine (IPV) induced higher GMTs of NA against poliovirusⅠand Ⅲ respectively. Among 182 children aged<5 years, the positive rate of NA against PVⅠ, Ⅱ, Ⅲ were 97.25%, 76.37% and 92.86%. There were statistical differences among the three types ( χ2=44.44, P=0.000). The rate of NA against PVⅡwas significantly lower than those ofⅠand Ⅲ (Ⅱ vs. I: χ2=34.65, P=0.000; Ⅱ vs. Ⅲ: χ2=18.99, P=0.000). And the GMTs of NA against PV Ⅰ, Ⅱ, Ⅲ were 1∶368.96、1∶23.06 and 1∶183.10, which were significantly different ( F=156.54, P=0.000). The GMT of PVⅠwas the highest, PV Ⅲ was the second, PVⅡ was lowest (pairwise comparison showed values of P all were 0.000). The analysis of general linear model showed that the interval between the date of last immunization and the date of sample collection would affect the GMTs of PVⅠand Ⅲ, different vaccine models would affect only the GMT of PV Ⅰ. The age maybe was a confounding factor. But no factors would affect the GMT of PV Ⅱ. Conclusions:After the vaccine switch, the level of NA against PVⅠand PV Ⅲ were still maintaining high level, but the level of PVⅡamong children<5 years was at relatively low level, which indicated that the surveillance should be strengthened.