Establishment of real-time fluorescent quantitative PCR for detection of torque teno virus types 7, 8 and 10 / 中华微生物学和免疫学杂志

Objective:To establish a real-time fluorescent quantitative PCR for the detection of torque teno virus types 7 (TTV7), 8 (TTV8) and 10 (TTV10) and analyze its performance in clinical sample detection.Methods:Specific primers were designed based on the gene sequences of TTV7, TTV8 and TTV10 in GenBank. Recombinant plasmids of pMD19-T-TTV7, pMD19-T-TTV8 and pMD19-T-TTV10 were constructed and used as positive standard control to establish a real-time fluorescent quantitative PCR based on FAM-Eclipse probe method. The specificity and sensitivity of the established method were evaluated. Moreover, it was validated in terms of clinical sample detection.Results:The standard curve equations of the real-time fluorescent quantitative PCR for detecting TTV7, TTV8 and TTV10 were y=-0.340 2 x+ 114.780 0 ( R2=0.998 8), y=-0.351 1 x+ 114.940 0 ( R2=0.995 3) and y=-0.348 9 x+ 115.020 0 ( R2=0.991 7), respectively, and there was no cross-reaction with other viruses. The detection sensitivity of the established method for TTV7, TTV8 and TTV10 were 108 copies/μl, 84 copies/μl and 98 copies/μl, and the positive detection rates in clinical pediatric serum samples were 10.9%, 2.1% and 4.3%, respectively. Conclusions:The established real-time fluorescent quantitative PCR for detection of TTV7, TTV8 and TTV10 was featured by strong specificity and high sensitivity, which could be used for rapid TTV detection in clinical serum samples.

Radial extracorporeal shock wave therapy for patients with subacromial impingement syndrome / 中华物理医学与康复杂志

Objective:To analyze the short-term therapeutic efficacy of radial extracorporeal shock wave therapy for patients with subacromial impingement syndrome.Methods:A total of 106 patients diagnosed as having subacromial impingement syndrome between October 2017 and April 2019 were randomized into a radial extracorporeal shock wave therapy (rESWT) group of 36, an exercise rehabilitation group of 35 and a conventional therapy group of 35. In addition to family exercise therapy, the rESWT group underwent 2000 to 2500 shots of extracorporeal shock wave therapy at 10 Hz and a pressure of 1.5-2.5 bar, once a week for four consecutive weeks. The exercise group was given range of motion exercises, joint control training and tendon movement training for 45 minutes, three times a week for four consecutive weeks. The conventional therapy group was treated with a laser apparatus and low-frequency electrotherapy, once a day, three times a week. Constant-Murle scores (CMSs) and the short form health survey (SF-36) were used to evaluate the clinical efficacy before and after 1 month of treatment.Results:Before the treatment there were no significant differences among the 3 groups in any of the measurements. After one month of treatment the average CMS pain score and total score of the exercise rehabilitation group were significantly better than the conventional therapy group′s averages. Moreover, the average body pain score, daily life ability, range of motion, muscle strength and total score of the rESWT group were all significantly better than the exercise and conventional therapy groups′ averages. In the SF-36 the average physical function, bodily pain, general health, and mental health scores of the rESWT groups were also significantly better than the other 2 groups′ averages.Conclusions:Radial extracorporeal shock wave therapy is superior to exercise therapy and conventional therapy for patients with subacromial impingement syndrome. It can restore shoulder joint function and improve the quality of life in one month.

Surface electromyography-based biofeedback for treating dysphagia after radiation therapy / 中华物理医学与康复杂志

Objective To observe the effect of surface electromyographic biofeedback (sEMG BFB) combined with routine swallow training in treating dysphagia among those with nasopharyngeal carcinoma after radiation therapy.Methods Fifty dysphagic patients with nasopharyngeal carcinoma after radiation therapy were randomly divided into a biofeedback training group and a routine treatment group,each of 25.Both groups were given routine training including orofacial function training,sensory irritation,behavioral swallowing training,and electrical stimulation.The biofeedback group was additionally given behavioral swallowing training based on sEMG BFB.Before and 4 weeks after the treatment,a videofluoroscopic swallowing study was performed to observe the opening of the upper esophageal sphincter (UES).The penetration aspiration scale (PAS) and the functional oral intake scale (FOIS) were used to evaluate the subjects' swallowing function.Results Before the treatment there were no significant differences between the two groups in terms of UES opening,average PAS score or average FOIS score.Everyone improved significantly after the treatment,but compared with the routine treatment group,UES opening was significantly better after the treatment,the average PAS score was lower and the average FOIS score was higher in the biofeedback training group.Conclusion sEMG BFB combined with routine swallowing training can improve the UES opening and swallowing ability of dysphagic patients with nasopharyngeal carcinoma after radiation therapy.

Establishment and verification of real-time fluorescent quantitative PCR detection system for ring virus 6 / 中华实验和临床病毒学杂志

Objective@#To establish a real-time quantitative PCR detection system for Torque teno virus (TTV) and verify the sensitivity and specificity of the detection system.@*Methods@#Primers and FAM-Eclipse probes were designed based on the TTV6 gene sequence registered in GenBank, and were to establish a real-time fluorescent quantitative PCR detecting way based on the FAM-Eclipse probe, the standard curve was constructed and sensitivity and specificity were analyzed.@*Results@#A quantitative PCR method for the specific detection of TTV6 were established that the standard curve equation was y=-3.0921x + 28.36, and the amplification efficiency and R2 were 99.6% and 1.000, respectively. The sensitivity of TTV6 was 1.0×10 copies/μl, and there was no cross-reactivity with other viruses. There was 1 case positive for TTV6 out of 56 throat swab samples from the patients with clinical respiratory infection.@*Conclusions@#The real-time fluorescent quantitative PCR for detecting TTV6 established by FAM-Eclipse probe had the advantages of high sensitivity and specificity. It provides an effective way for detection and quantification of viral content of TTV6 in clinical specimens.

Changes of endogenous small interfering RNA in H1-HeLa cells induced by human rhinovirus 16 infection / 中华实验和临床病毒学杂志

Objective@#To observe the changes of endogenous small interfering RNA (siRNA) in H1-Hela cells infected with human rhinovirus 16 (HRV 16).@*Methods@#To determine whether HRV16 infection induced the changes of siRNA, H1-HeLa cells were infected with HRV16 for 12 h, 24 h and 36 h, siRNAs were detected by high-throughput sequencing, second-generation sequencing) and qRT-PCR.@*Results@#The result showed that siRNA was generated differently at different time points post-infection, among which novel_sir907 and novel_sir1950 decreased at three time points. Further validation by qRT-PCR showed that novel_sir907 decreased at 12 h, 24 h and 36 h post-infection compared with the cell control, but novel_sir1950 increased at 12 h then decreased at 24 h and 36 h.@*Conclusions@#HRV16 infection induces changes endogenous siRNAs.

A preliminary study on thapsigargin promoting coxsackievirus B3 replication through activating transcription factor 6 pathway / 中华实验和临床病毒学杂志

Objective@#To investigate the effect of thapsigargin (TG) which can induce endoplasmic reticulum stress (ERS) on the replication of coxsackievirus B 3 (CV-B3).@*Methods@#After 10 MOI CV-B3 infected HeLa cells were exposed 0.25 μmol/L TG for 3 h, 6 h and 9 h, virus RNA of HeLa cells were extracted and viral replication was evaluated by real time PCR. After 0.25 μmol/L、0.08 μmol/L and 0.025 μmol/L TG exposed, the plaque of CV-B3 was used to confirm further replication of CV-B3. To verify TG induced ERS through three signal pathway, one of among PERK, ATF6 and IRE1 inhibitors GSK2656157, AEBSF and STF-083010, and 0.25 μmol/L TG were used in HeLa cells infected with 10 MOI CV-B3, replication of CV-B3 was evaluated by qRT-PCR.@*Results@#The stimulation of TG did not induce increase of virus replication after post-infection 3 h. However, TG induced replication of virus to increase 2.5 times after post-infection 6 h and 158.6 times after post-infection 9 h. And, the area of viral plaque was significantly increased. ATF6 inhibitors AEBSF significantly inhibited promotion of virus replication from TG.@*Conclusions@#TG can promote the replication of CV-B3 through ATF6 signal pathway.

A autophagy induced by non-stuctural protein 2B of human rhinovirus / 中华实验和临床病毒学杂志

Objective@#To study the intracellular location and autophagosome production of rhinovirus 16 2B protein using miniSOG labeling technique.@*Methods@#2B was fused with miniSOG and flag tags to construct pcDNA3.1-2B-miniSOG-flag plasmid, which was used to transfect HEK293 cells, LC3 protein was detected by western blot. The transfected cells were fixed, stained with DAB through the photooxidation activity of miniSOG, and used to prepare ultrathin sections. Localization of 2B-miniSOG protein in cells and ultrastructural changes of cells were observed under electron microscope.@*Results@#2B-miniSOG protein glows green under a fluorescence microscopy. Green flourescence coold be observed in the cells expressing 2B-miniSOG protein.LC-II protein increased in the cells transfected with pcDNA3.1-2B-miniSOG-flag. Under electron microscopy it was observed that 2B-miniSOG protein was located in the mitochondria, and a large number of vesicular structures appeared in the cytoplasm. Both autophagosomes and autophagic lysosomes can be observed.@*Conclusions@#Non-structural protein 2B of HRV16 can induce autophagy.

Analysis on global Rhinovirus research advances / 中华实验和临床病毒学杂志

Objective@#To explore the research status of rhinovirus (RV) through analysis of rhinovirus literature using GoPubMed.@*Methods@#"Rhinovirus" was used as the major subject word and the rhinovirus literature was collected at PubMed database (from Jan 1, 1970 to April 16, 2018). The high frequency subject words of rhinovirus related literature and the distribution of countries, cities, and journals were analyzed through a bibliometrical analysis method .@*Results@#A total of 5 367 reports were retrieved from PubMed. The quantity of rhinovirus papers increased overall year by year. The highest number of papers were mainly published in developed countries. The highest number of papers on RV were mainly published in J Virol among all journals related with rhinovirus and Tyrrell D published the highest number of papers in all authors contributed to articles on rhinovirus. The rhinovirus, human, virus, respiratory tract infection were the high frequency subject words in the rhinovirus research.@*Conclusions@#Rhinovirus research is becoming one of research hotspots according to the statistical analysis of the research literature on rhinoviruses by GoPubMed.

Establishment of a real-time PCR method to identify Ekpoma virus gene in blood sample of a returnee from Angola / 中华实验和临床病毒学杂志

Objective@#To establish quantitative real-time PCR (qPCR) method based on Taqman probe for detecting Ekpoma virus (EKV).@*Methods@#According to the conserved region of gene in EKV genome from GenBank, primers and probe for qPCR were designed. Validity and sensitivity were evaluated in this study. Both whole blood and serum of a returnee from Angola were tested by the established EKV-1 and EKV-2 qPCR method .@*Results@#Sensitivity of EKV-1 and EKV-2 qPCR method was respectively 41 copies/μl and 70 copies/μl. Coefficient of variance (CV) was respectively 1.27%, 0.20%, 0.82%; 2.12%, 1.74%, and 1.40%. EKV-2 gene was detected in both whole blood and serum of a returnee from Angola.@*Conclusions@#The first EKV-2 gene was confirmed in both whole blood and serum of a returnee from Angola by real-time RT-PCR..

TNF-β genetic variant contributes to the risk of non-small cell lung cancer / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the association of TNF-β 252A >G variant with the risk of non-small cell lung cancer (NSCLC).</p><p><b>METHODS</b>Total 956 patients with NSCLC were recruited between January 2000 and December 2008 at Cancer Hospital, Chinese Academy of Medical Science as the case group, and 994 frequency-matched controls were randomly selected from a pool of cancer-free subjects recruited from a nutritional group. All the participants were unrelated Han Chinese. There were no age, gender restrictions. Smoking status of the subjects was surveyed. Informed consent was obtained and 3 ml peripheral blood was collected from each subject. All samples were genotyped by polymerase chain reaction-restriction fragment length polymorphism method (PCR-RFLP). The OR and 95%CI were estimated by logistic regression to evaluate the relationship between TNF-β 252 A/G variant and the risk of lung cancer.</p><p><b>RESULTS</b>The frequencies of TNF-β 252 AA, AG and GG genotype were 30.9% (307/994) , 47.4% (471/994) and 21.7% (216/994) in lung cancer cases and 35.7% (341/956) , 48.1% (460/956) and 16.2% (155/956) in controls. Logistic regression analysis revealed that TNF-β 252 GG genotype contributed to a decreased risk of developing NSCLC (OR = 0.64, 95%CI: 0.49-0.83) compared with AA genotype. When stratified by smoking status, the individuals with 252 GG genotype had a significant increased risk of NSCLC (OR = 1.54, 95%CI:1.00-2.39) among smokers; which was less than those with AA genotype among smokers (OR = 2.88, 95%CI:1.91-2.24). When further stratified by smoking index, individuals with 252 GG genotype had a significant decreased risk of NSCLC among heavy smokers with OR (95%CI) of 2.24 (1.33-3.74), which was less than those with AA genotype (OR = 4.62, 95%CI:2.88-7.41).</p><p><b>CONCLUSION</b>TNF-β genetic variant may interact with environment factor to contribute to the susceptibility to NSCLC.</p>

The relationship between Gly587Arg variant of COX-2 gene and the primary liver cancer / 中国综合临床

Objective To explore the association of COX-2 Gly587Arg polymorphism with the risk of primary liver cancer.Methods Two hundred and seventy patients with primary liver cancer and 540 health people were selected as our subjects.DNA were extracted from peripheral blood lymphocytes,and genotypes were measured by polymerase chain reaction-restriction fragment length polymorphism method.Odds ratios(OR) and 95％ confidence intervals(CI) were estimated by logistic regression.Results Two kinds of genotype (587Gly/ Gly and Gly/Arg) were found in all participants.No one carried 587Arg/Arg genotype.Among primary liver cancer patients,91.5％ (247/270,) 8.5％ (23/270) of individuals carried 587Gly/Arg and Gly/Arg genotype,which was significantly higher than that of controls (96.5％ (521/540,) 3.5％ (19/540)).Multivariate Logistic regression analysis showed that individual carried 587Gly/Arg genotype had an increased risk of developing primary liver cancer (OR =2.56,95％ CI =1.37-4.79,P =0.003) compared with 587Gly/Gly carriers.Conclusion COX-2 Gly587Arg polymorphism is a risk factor for primary liver cancer in Han.