Objective:A recombinant adenoviral vector vaccine based on non-replicating human adenovirus type 5 (Ad5), encoding the conserved domain of respiratory syncytial virus G protein (RSV-G) was constructed. The immunogenicity and protective efficacy of this vaccine were subsequently evaluated in mice.Methods:The recombinant Ad5 vector plasmid (Ad5-Gbcc-Gacc) was constructed by inserted conserved domains of RSV A and RSV B. The recombinant adenovirus Ad5-Gbcc-Gacc was rescued in HEK293A cells. The genome of virus Ad5-Gbcc-Gacc was identified by multi-enzyme digestion, and the expression of Ad5-Gbcc-Gacc was verified by Western blot. Recombinant adenovirus was used to immunize BALB/c mice via intramuscular injection with signal dose, and then challenged with RSV Long strain at week 6. The levels of G specific IgG and antibody subtypes in serum were detected by enzyme-linked immunosorbent assay, the level of neutralizing antibodies was determined by micro-neutralization assay. After challenge, the mice′s weight was recorded daily, the copies of RSV virus in the lung and nasal tissues were detected. Pathological changes in lung tissue were also examined.Results:Western blot and multi-enzyme digestion identification confirmed the successful rescue of the recombinant adenovirus. Ad5-Gbcc-Gacc elicit high titers of specific IgG, robust neutralizing antibodies, and a balanced Th1/Th2 immune response in mice. In comparison to unimmunized controls, mice immunized with Ad5-Gbcc-Gacc reduced the viral copies in both lung and nasal tissue, and exhibited only minimal pathological damage of lung tissue following RSV challenge. In conclusion, Ad5-Gbcc-Gacc induced robust immunogenicity and offers protective effects against RSV infection in murine models.Conclusions:Ad5-Gbcc-Gacc induce robust immunogenicity and can protect mice from RSV challenge, which lays a foundation for further development of RSV vaccine based on G protein.

[Purpose]To investigate the trends of incidence and age at onset of uterine corpus can-cer in Jiangsu cancer registration areas from 2009 to 2019.[Methods]The continuous monitoring data of uterine corpus cancer from 2009 to 2019 were collected from 16 cancer registries in Jiang-su Province.The crude incidence rate,the age-standardized incidence rate by Chinese standard population(ASIRC),crude and adjusted mean age,and standardized age-specific incidence composition were calculated.The average annual percentage change(AAPC)were analyzed by the Joinpoint regression model.The linear regression model was used to analyze the relationship be-tween mean age at onset and year.The standardized age-specific incidence composition in 2009 and 2019 were compared.[Results]The ASIRC of uterine corpus cancer in all registration areas and in rural areas of Jiangsu Province showed upward trends with AAPC of 1.78％and 2.38％,re-spectively(P＜0.05),but not showed in the urban areas(AAPC=1.30％,P＞0.05).The crude mean age at onset increased from 56.48 years old in 2009 to 58.26 years old in 2019 with an average annual growth of 0.173 years old(P=0.001).After the population structure standardized,the trends disappeared in all registration areas.[Conclusion]From 2009 to 2019,the standardized incidence rates of uterine corpus cancer were on rise in Jiangsu cancer registration areas,especially in the age group of 50 to 59 years old.

Enterotoxigenic Escherichia coli（ETEC）infection can induce watery diarrhea，leading to dehydration，electrolyte disturbance，and even death in severe cases. Recombinant B subunit/inactivated whole-cell cholera（rBS/WC）vaccine is effective in preventing ETEC infectious diarrhea. On the basis of the latest evidence on etiology and epidemiology of ETEC，as well as the effectiveness，safety，and health economics of rBS/WC vaccine，National Clinical Research Center for Child Health（The Children’s Hospital，Zhejiang University School of Medicine）and Zhejiang Provincial Center for Disease Control and Prevention invited experts to develop expert consensus on rBS/WC vaccine in prevention of ETEC infectious diarrhea. It aims to provide the clinicians and vaccination professionals with guidelines on using rBS/WC vaccine to reduce the incidence of ETEC infectious diarrhea.

Objective:To understand the prevalence of human parechovirus (HPeV) in neonates of Hunan Provincial People’s Hospital, and analyze genetic evolutionary characteristics.Methods:From June to September 2021, fecal samples of inpatient neonates were collected in Hunan Provincial People′s Hospital. TaqMan real-time qPCR and RT-PCR were used for HPeV screening and genotyping. High-throughput sequencing and PCR were used to obtain whole genomes. Phylogenetic analysis was performed after sequencing.Results:A total of 123 fecal samples of neonates were collected, of which 22 were HPeV positive, with 17.89% positive rate. All the strains belonged to the HPeV-1 genotype. One full-length genomic sequence of 7 269 bp were obtained, and provisionally named Hunan/HPeV/2021, which has the highest nucleotide identity with known HPeV-1 genotype, with 86.6%-91.9% nucleotide identity. The nucleotide and amino acid identity of open reading frame (ORF) with known similar sequences were 90.3%-92.6% and 97.3%-98.3%, respectively. The phylogenetic analysis showed that Hunan/HPeV/2021 belongs to the HPeV-1 genotype, which is clustered into the same clade as the popular HPeV-1 strains in China.Conclusions:HPeV has a high prevalence in inpatient neonates of Hunan Provincial People’s Hospital and belong to the HPeV-1 genotype.

Objective:To optimize and evaluate the pretreatment method for high-throughput sequencing of fecal and tissue samples to improve the sensitivity of high-throughput sequencing in the study of virome.Methods:For fecal samples, five virus-positive samples that have been detoxified from feces were selected, mixed as the simulated samples, and filters made of different materials were used, different processing times were set for nucleases, and different kits were used to extract nucleic acid. For tissue samples, two virus-positive samples that have been detected in animal tissues, filter and nuclease treatment were used, and different extraction method were used to extract nucleic acid. TaqMan real-time PCR was used to quantitatively evaluate the impact of each treatment on the virus and the advantages and disadvantages were compared. We used the SYBR Green real-time PCR quantitative method to evaluate the removal effect of the above method on bacteria 16S rRNA and host 12S rRNA genome.Results:For fecal samples, the 0.22 μm PES filter showed a better filtering effect, and the PVDF material filter reduces the sample volume; 2 h nuclease digestion was better than 1 h digestion to remove bacteria, and the virus loss was less; the use of RPMK kits can effectively reduce bacteria, but the effect of extracting some viruses was poor, and the MVSK kit has a better effect of extracting viral nucleic acid. For tissue samples, 0.22 μm PES filter filtration, nuclease digestion for 1 h and VNAEK II kit extraction of nucleic acid were the best, Trizol LS combined with the RPMK method was better for gDNA removal, but the virus loss was larger. The virus loss of the whole process of the pretreatment method of feces and tissue samples was (1.7-3.0) Ct and (1.6-2.5) Ct, respectively.Conclusions:The optimal method for fecal samples was to filter with a PES filter, then digest with nuclease for 2 hours, and then extract nucleic acids using the MVSK kit; the optimal method for tissue samples was to filter with a PES filter, then perform 1 h nuclease digestion, and then use VNAEK II kit to extract nucleic acids.

Objective To systematically evaluate the efficacy and safety of 125I seed implantation for treatment of advanced pancreatic cancer.Methods An electronic literatuire search was performed about randomized controlled trials(RCTs) of 125I implantation for treamtent of advanced pancreatic cancer in CNKI,Wanfang Data,CBM,Cochrane Library,PubMed and Embase (from the date of building the database to November 2016).Two investigators independently screened literature,extracted data and assessed the risk bias of included studies,and the Meta-analysis was performed by using Revman 5.3software.Results There were 12 RCTs (n =689) included.Meta-analysis showed that the objective respond rate(ORR) (OR =3.24,95％CI2.33-4.52,P＜0.001),the 6-month survival rate(OR =3.61,95％ CI 1.53-8.52,P =0.003),the 12-month survival rate(OR =4.80,95％ CI 2.40-9.57,P ＜ 0.001) and the relief rate of pain were higher than those in the control group.However,there were no significant differences between both groups in the 2-year survival rate and the adverse reaction rate,which were (OR=2.36,95％ CI 0.47-11.74,P =0.29) and (OR =4.94,95％ CI 1.05-23.23,P =0.04),respectively.Conclusions The limited current evidence showed that 125I implantation for treatment of advanced pancreatic cancer is effective and safety.125I implantation can improve the ORR,short-time survival rate and pain relief rate.In addition,there was no significant increase in the incidence of related adverse events except for seed malposition.Although the quality and quantity of evidences is limited,it merits further study to provide high quality evidences.

Objective To investigate the efficacy and side effects of radioactive seed implantation in the treatment of non-surgical early stage non-small cell lung cancer (NSCLC) based on CT guidance combined with template.Methods Twenty-one patients with inoperable T1 2N0M0 NSCLC who underwent CT-guided radioactive seed implantation therapy were retrospectively analyzed from December 2010 to October 2016 in the Second Hospital of Tianjin Medical University.All patients were diagnosed by histopathology.All seeds,with the activity of 18.5-29.6 MBq and prescription dose of 120-160 Gy,were completed in an operation of the radioactive seed implantation.The preoperative and postoperative TPS treatment plans and quality verification were corducted.In addition,the local control rate of tumors,overall survival (OS),progression free survival time (PFS),satisfaction rate of dose validation and adverse reactions were evaluated.Results The median follow-up was 25.1 months (range 4.4-72.7months).The local control rate of primary tumor in 1-,2-and 3-year was 100％,95.2％ and 95.2％,respectively.Of all patients,the median OS was 48 months with the median PFS 43.4 months.In particular,the 1-,2-and 3-year survival rate was 100％,91.7％ and 72.9％,respectively.Moreover,the rate of 3-year PFS was 70.2％ and the satisfactory rate of postoperative quahty verification was 100％.The treatment-related adverse events included pneumothorax,bronchial hemorrhage,pleural effusion,cough,pulmonary fibrosis and seed shifts.In all,7 (33.3％) patients had grade 1 adverse events and 4 (19％) patients with grade 2,but no grade 3 adverse event.Conclusions CT and template-guided radioactive seed implantation in NSCLC with T1 2 N0 M0 has a high tumor local control rate and low treatment-related adverse reactions,suggesting that it might provide an alternative way for the treatment of inoperable early stage NSCLC.

Objective To evaluate the clinical effectiveness and safety of stents loaded with 125I seeds compared to conventional stents.Methods Literatures were searched in PubMed,EMbase,Cochrane Library,CBM,CNKI,Wanfang Data and other electronic databases from inception to November 2016.Two reviewers independently screened the literature according to the inclusion and exclusion criteria,extracted data and assessed quality of the included studies independently.Meta-analyses were performed using RevMan 5.3.Results A total of five RCTs and 14 CCTs involving 1 211 patients were included.The mean survival time of the 125I stent group was significantly higher than that of the control group [mean difference =4.11,95％ CI (2.16-6.07)P ＜0.001].The incidence of restenosis after 3:The available data showed that the incidence of re-staging of 125I stent in the treatment group was lower than that of the normal stent group [RR =0.23,95％ CI(0.12-0.62),P =0.002].Postoperative bleeding [RR =0.80,95％CI (0.52-1.23),P=0.30];Postoperative pain[RR=1.06,95％CI(90.88-1.27),P=0.55];postoperative stent shift [RR =0.53,95％ CI(0.27-1.05),P =0.07].The difference of incidence of complications was not statistically significant.There was no difference in the incidence of complications between the two groups.Conclusions The available data suggest that 125I stent is superior to common stent in the treatment of advanced esophageal cancer.There are no differences found in the incidence of complications between 125I stent and conventional stent.However,due to the limited quality of the included studies,more high-quality and multicenter-based studies are needed to verify the above conclusion.

Objective To investigate thc clinical effect of midazolam in CT-guided 125I seed implantation.Methods Totally 86 patients who underwent CT-guided radioactive 125I seed implantation were collected from December 2015 to February 2017 in the Second Hospital of Tianjin Medical University and randomly divided into two groups:the test group and the control group.The test group was given intravenous infusion of midazolam and local anesthesia of lidocaine while the control group were given the same amount of saline and lidocaine.The changes of heart rate (HR),mean arterial pressure (MAP),respiration (RR) and oxygen saturation (SpO2) were observed and compared between the two groups at 15 min before the midazolam(T0),15 min after the midazolam (T1),15 min after lidocaine local anesthesia (T2),the moments the lesion was punctured (T3),and after implantation of the seeds (T4) and the end of the operation (T5),respectively.In addition,the body reaction,operation time,the amount of lidocaine,the effect of analgesic and adverse reactions were also observed.Results The levels of HR,MAP,RR and SpO2 in test group were significantly lower than those in control group (t =38.9,31.0,14.1,2.4,P ＜0.05),but there was no significant difference at T1,T2,T3,T4 and T5(P ＞0.05).In control group,HR,MAP and RR were significantly higher than those before operation (t =-23.6,-18.0,-9.5,P＜0.05).The difference ofHR,MAP,RR and SpO2 among T1,T2,T3,T4 and T5 was statistically significant (F =997.3,833.7,120.1,77.5,P ＜ 0.05).In addition,the incidence of adverse reactions was both less in the two groups,and the incidence of response of patients,operation time,the amount of lidocaine and other indicators and analgesic effect in the test group were superior to the control group (-15.4,9.2,-56.3,P ＜ 0.05).Conclusions Midazolam in CT-guided 125I particle implantation can improve the patient's tolerance and shorten the operation time,guarantec security.

Objective To investigate the human papillomavirus 16/18 (HPV16/18) infective status and its relationship with the expression of p16,CK17,Ki-67 in immature squamous metaplasia(IM),cervical intraepithelial neoplasia (CIN) and cervical carcinomas (CC).Methods In situ hybridization (ISH) and immunohistochemistry were applied to detect the expression of HPV16/18,p16,CK17,Ki-67 on tissues from 30 IM,60 CIN and 30 CC.Results The expression rates of HPV16/18 in LSIL,HSIL and CC were 30 ％ (9/30),60 ％ (18/30) and 77 ％ (23/30),higher than that in IM [6.7 ％ (2/30)].HPV16/18 was positively expressed with the progression of cervical squamous epithelial lesions.The expression rates of CK17 in IM was 90 ％ (27/30),higher than that in LSIL,HSIL and CC [17 ％ (5/30),10 ％ (3/30),6.7 ％ (2/30)].The expression rates of p16,Ki-67 were 83 ％ (25/30) and 87 ％ (26/30) in LSIL,90 ％ (27/30) and 93 ％ (29/30) in HSIL,97 ％ (29/30) and 97 ％ (29/30) in CC,higher than that in IM [13 ％ (4/30) and 10 ％ (3/30)].The expression of p16,Ki-67 were particularly seen in HPV16/18-positive HSIL and CC,and the correlation were observed.Conclusion HPV16/18 infection is highly associated with the degree of squamous intraepithelial neoplasia and carcinomas of cervix and upregulated p16 and Ki-67,which suggested that HPV16/18 maybe cause mutation of p16 gene.With the progression of cervical squamous epithelial lesions,positive expression rates of CK17 were decreased compared with increased of HPV16/18,p16,Ki-67.Combined detection of HPV16/18,p16,Ki-67 is helpful for diagnosis and classification of cervical squamous epithelial lesions.