1.Detection of G9P[4]rotavirus in the surveillance of sporadic viral diarrhea in Jiangsu province,China,in 2023
Chuchu LI ; Jing AI ; Yuanfang QIN ; Liguo ZHU ; Shenjiao WANG ; Changjun BAO ; Jianguang FU
Chinese Journal of Experimental and Clinical Virology 2025;39(5):611-616
Objective:To investigate the G/P genotypes of group A rotavirus(RVA)in the 2023 sentinel surveillance in Jiangsu Province,and to conduct a molecular characterization analysis of the whole-genome sequences of four G9P[4]genotype RVA strains identified during surveillance.Methods:A total of 212 RVA-positive specimens collected from the surveillance system in 2023 were subjected to G/P genotyping using multiplex nested RT-PCR. Whole-genome sequencing was performed on six G9P[4]strains. The resulting complete genome sequences were preliminarily genotyped using BLAST,followed by comprehensive molecular characterization analyses utilizing BioEdit 7.0.5,MAFFT,MEGA 7.0,and iTOL software.Results:The overall RVA positivity rate was 6.22%. The predominant G/P combination in both outpatient and inpatient settings was G8P[8]. Among the six G9P[4]strains,four were successfully sequenced. All four exhibited the genotype constellation G9-P[4]-I2-R2-C2-M2-A2-N1-T2-E2-H2. While the NSP2 gene belonged to the N1 genotype,all other genes corresponded to the DS-1-like genogroup. Phylogenetically,the four Jiangsu G9P[4]strains clustered within Lineage V of the VP7 gene and formed a distinct minor subclade within the N1 branch of the NSP2 gene. Unique amino acid substitutions were identified at multiple VP7 neutralization antigenic epitope sites when compared to vaccine strains.Conclusions:The predominant circulating RVA strain in Jiangsu province during 2023 was G8P[8]. Concurrently,the relatively uncommon G9P[4]-N1 strain was detected. This strain exhibited significant amino acid differences at key epitopes compared to vaccine strains. Enhancing the proportion of whole-genome sequencing in RVA surveillance is warranted to obtain more detailed genetic information,thereby providing crucial data to support future vaccine development and optimization strategies.
2.Correlation between type 2 diabetes and hepatitis B virus infection
Wu LIU ; Tianshuo ZHAO ; Liguo YANG ; Genxia GAO ; Wenli MIAO ; Tongxia ZENG ; Yonglan CHEN ; Lina ZHANG ; Xiaoyan CHE ; Lihong WANG ; Junhan ZHANG ; Bingfeng HAN ; Fuqiang CUI
Chinese Journal of Epidemiology 2025;46(5):833-839
Objective:To analyze the latent prevalence of hepatitis B and type 2 diabetes and their correlation through an observational study.Methods:This study used a case-control design. The cases with diabetes were recruited through the diabetes management system and village doctors, while the controls without diabetes were screened from volunteers recruited by village health clinics. Capillary blood samples were collected from the study participants for the measurement of real-time blood glucose level, and venous blood samples were taken from them for the detections of HBV serological markers. Firth logistic regression model was used to fit the relationship between HBsAg positive status and diabetes status.Results:The study included 1 218 diabetes patients, 62 patients with impaired fasting glucose and 491 cases without diabetes. In the cases without diagnosis of diabetes, 11.15% had impaired fasting blood glucose and 4.43% had diabetes. Among those who reported no or unknown diagnosis of hepatitis B, 1.73% were positive for HBsAg, while 18.80% were positive for both HBV core antibody and surface antibody, indicating latent infection of hepatitis B virus. In the non-diabetes group, 0.81% reported hepatitis B history, and in the diabetes group, 2.76% reported hepatitis B history. After adjustment, the HBsAg positive rate was higher in the diabetes group ( OR=2.90, 95% CI: 1.21-6.91). Conclusions:Both diabetes and hepatitis B exhibited a high degree of latent prevalence. The HBsAg positive rate was significantly higher in those with diabetes than in those without diabetes, indicating a potential correlation. These findings highlighted the importance of strengthened screening and management of comorbidities.
3.Study on the immunogenicity and persistence of different types of hepatitis B vaccines in diabetic patients
Wu LIU ; Tianshuo ZHAO ; Liguo YANG ; Genxia GAO ; Wenli MIAO ; Xiaoyan CHE ; Lihong WANG ; Junhan ZHANG ; Fuqiang CUI
Chinese Journal of Epidemiology 2025;46(8):1409-1416
Objective:Exploration of the immunogenicity and persistence of three different immunization regimens of hepatitis B vaccines in diabetic patients.Methods:Participants with diabetes and non-diabetic individuals were recruited from study sites and assigned to different vaccination regimens: the diabetic group (①D60Yeast0-1: received 60 μg Saccharomyces cerevisiae-derived recombinant HBV vaccine on a 0-1-month schedule; ②D20Yeast0-1-6: received 20 μg Saccharomyces cerevisiae-derived recombinant HBV vaccine on a 0-1-6-month schedule; ③D20CHO0-1-6: received 20 μg Chinese hamster ovary (CHO) cell-derived recombinant HBV vaccine on a 0-1-6-month schedule) and the non-diabetic group (ND20Yeast0-1-6: non-diabetic individuals received 20 μg Saccharomyces cerevisiae-derived recombinant HBV vaccine on a 0-1-6-month schedule). Venous blood samples were collected at 1,12, and 48 months post-full vaccination to measure anti-HBs levels. Differences in immunogenicity between diabetic and non-diabetic groups, as well as among diabetic subgroups, were analyzed.Results:This study enrolled a total of 564 subjects. In the D20CHO0-1-6 group, the seroconversion rate decreased from 90.72% (95% CI: 84.84%-96.60%) at 1 month to 74.23% (95% CI: 65.37%-83.08%) at 48 months, and the antibody geometric mean concentration (GMC) decreased from 676.08 (95% CI: 389.05- 1 148.20) mIU/ml at 1 month to 33.11 (95% CI: 23.44-46.77) mIU/ml at 48 months. In the D20Yeast0-1-6 group, the seroconversion rate declined from 93.81% (95% CI: 89.29%-98.32%) at 1 month to 63.72% (95% CI: 54.71%-72.72%) at 48 months, with antibody GMC dropping from 630.96 (95% CI: 407.40-954.99) mIU/ml to 25.70 (95% CI: 17.78-38.02) mIU/ml over the same period. For the D60Yeast0-1 group, seroconversion rate fell from 82.03% (95% CI: 75.29%-88.77%) to 56.25% (95% CI: 47.54%-64.96%), and antibody GMC decreased from 81.28 (95% CI: 51.29-128.82) mIU/ml to 15.49 (95% CI: 11.75-20.89) mIU/ml between 1 and 48 months. The ND20Yeast0-1-6 group (non-diabetic control) exhibited a higher initial seroconversion rate of 97.56% (95% CI: 94.80%- 100.00%) at 1 month, but it still declined to 76.42% (95% CI: 68.82%-84.03%) at 48 months, with antibody GMC decreasing from 1 318.30 (95% CI: 912.01- 1 905.50) mIU/ml to 34.67 (95% CI: 25.12-47.86) mIU/ml. Multivariate analysis on factors influencing the GMC of antibodies revealed statistically significant differences in antibody GMC between the D20Yeast0-1-6 group and ND20Yeast0-1-6 group at 12 months (a OR=0.73, 95% CI: 0.58-0.93) and 48 months (a OR=0.79, 95% CI: 0.63-0.99) post-vaccination (all P<0.05). As for the diabetic population, when compared with the D20Yeast0-1-6 group, the D60Yeast0-1 group also showed statistically significant differences in antibody GMC at 12 months (a OR=0.57, 95% CI: 0.44-0.74) and 48 months (a OR=0.60, 95% CI: 0.47-0.76)(all P<0.05). Conclusions:The seroconversion rate and antibody GMC gradually decreased over time (1, 12, and 48 months) in the four groups. Diabetic patients showed poor immunogenicity and persistence to hepatitis B vaccines. The immunogenicity and persistence of hepatitis B vaccination in diabetic patients were associated with vaccine type, antigen dose, and vaccination regimen. The CHO cell-recombinant hepatitis B vaccine demonstrated better performance in terms of immunogenicity and persistence among the diabetic population.
4.Detection of G9P[4]rotavirus in the surveillance of sporadic viral diarrhea in Jiangsu province,China,in 2023
Chuchu LI ; Jing AI ; Yuanfang QIN ; Liguo ZHU ; Shenjiao WANG ; Changjun BAO ; Jianguang FU
Chinese Journal of Experimental and Clinical Virology 2025;39(5):611-616
Objective:To investigate the G/P genotypes of group A rotavirus(RVA)in the 2023 sentinel surveillance in Jiangsu Province,and to conduct a molecular characterization analysis of the whole-genome sequences of four G9P[4]genotype RVA strains identified during surveillance.Methods:A total of 212 RVA-positive specimens collected from the surveillance system in 2023 were subjected to G/P genotyping using multiplex nested RT-PCR. Whole-genome sequencing was performed on six G9P[4]strains. The resulting complete genome sequences were preliminarily genotyped using BLAST,followed by comprehensive molecular characterization analyses utilizing BioEdit 7.0.5,MAFFT,MEGA 7.0,and iTOL software.Results:The overall RVA positivity rate was 6.22%. The predominant G/P combination in both outpatient and inpatient settings was G8P[8]. Among the six G9P[4]strains,four were successfully sequenced. All four exhibited the genotype constellation G9-P[4]-I2-R2-C2-M2-A2-N1-T2-E2-H2. While the NSP2 gene belonged to the N1 genotype,all other genes corresponded to the DS-1-like genogroup. Phylogenetically,the four Jiangsu G9P[4]strains clustered within Lineage V of the VP7 gene and formed a distinct minor subclade within the N1 branch of the NSP2 gene. Unique amino acid substitutions were identified at multiple VP7 neutralization antigenic epitope sites when compared to vaccine strains.Conclusions:The predominant circulating RVA strain in Jiangsu province during 2023 was G8P[8]. Concurrently,the relatively uncommon G9P[4]-N1 strain was detected. This strain exhibited significant amino acid differences at key epitopes compared to vaccine strains. Enhancing the proportion of whole-genome sequencing in RVA surveillance is warranted to obtain more detailed genetic information,thereby providing crucial data to support future vaccine development and optimization strategies.
5.Effect of practicing Traditional Chinese Medicine guidance on health status in weightless conditions
Xiaorui WU ; Haijun HAN ; Junlian LIU ; Quanchun FAN ; Liguo GUO ; Yu LIU ; Jiaping WANG ; Jihong SHEN ; Xining CHEN
Space Medicine & Medical Engineering 2025;36(4):385-388
Objective To study the effect of practicing traditional Chinese medicine(TCM)guidance on the health in weightless conditions based on the examination results of the TCM diagnostic instrument.Methods 6 subjects in weightless conditions practiced TCM guidance for 14 consecutive days,once a day for 8 minutes each time.The Digital TCM Diagnostic Instrument and TCM Syndrome Diagnosis Scale were used to collect information before and after their practice of TCM guidance through diagnostic methods of observation,palpation and inquiry according to TCM theory.The data such as the degree of virtual-reality,pulse rate,and characteristic parameters of pulse diagrams h3/h1,h4/h1,t1,h1/t1 were processed and analyzed,and the changes before and after the practice were compared.Results Compared with before the practice,the degree of deficiency syndrome was significantly reduced after practicing TCM guidance(P<0.05),the pulse rate was significantly decreased(P<0.01),and the pulse diagnosis parameters h3/h1 and t1 were significantly decreased(P<0.05),while h4/h1 and h1/t1 showed no significant changes.Conclusion Practicing TCM guidance in weightless conditions is helpful for enhance cardiovascular regulatory function,correcting symptom bias,adjusting the body's balance state,and has a positive effect on health.
6.Short-term outcome study on cervical deep lymph node-venous anastomosis technique in the treatment of Alzheimer’s disease
Cheng GAN ; Zhengdong KONG ; Xiaoye RAN ; Shudong QIAO ; Yixin ZHANG ; Lu YUE ; Yingjie WANG ; Hui BI ; Dong YANG ; Hongtong MA ; Yuan CHEN ; Hongli CHAI ; Ying JIA ; Chenhao MA ; Zixiang CHEN ; Ke LI ; Miao WANG ; Liguo XUE ; Siwen ZHAO ; Ke WEN ; Lin YIN ; Bo DING ; Shan ZHU ; Yuanbo LIU ; Mengqing ZANG
Chinese Journal of Plastic Surgery 2025;41(2):130-143
Objective:To explore the short-term clinical effects of deep cervical lymph node-venous anastomosis in the treatment of Alzheimer’s disease (AD).Methods:A prospective exploratory study was conducted on the treatment of AD patients using the cervical deep lymph node-venous anastomosis technique in Scar and Wound Treatment Department, Plastic Surgery Hospital, Chinese Academy of Medical Sciences, from September to October 2024. The patients underwent high-frequency ultrasound to locate deep cervical lymph nodes and the external jugular vein. Under general anesthesia, bilateral deep cervical lymph node-venous anastomoses were performed. Indocyanine green (ICG) lymphography was conducted via subcutaneous injection behind the ear to visualize lymph nodes in levels Ⅱ and Ⅲ. After making a skin incision along the posterior margin of the sternocleidomastoid muscle, the external jugular vein, internal jugular veins, and associated lymph nodes were exposed. Adjacent veins were selected for anastomosis of lymph node. Using microsurgical techniques, end-to-side or end-to-end anastomosis was completed for lymph nodes in levels Ⅱ and Ⅲ. Preoperative assessments included the mini-mental state examination (MMSE, a higher score indicates better cognitive function), Alzheimer’s disease assessment scale-cognitive subscale (ADAS-Cog, a higher score indicates greater impairment of cognitive function), Alzheimer’s disease cooperative study scale for activities of daily living (ADCS-ADL, a higher score indicates better ability to perform daily activity), and neuropsychiatric inventory (NPI, a higher score indicates more severe behavioral and emotional symptom). Postoperative follow-up included the same scales to observe changes in cognitive function, activities of daily living, and emotional communication.Results:Four patients (1 male, 3 females, aged 58-79 years) with AD were included. All were diagnosed based on cerebrospinal fluid biomarkers. All patients successfully underwent bilateral deep cervical lymph node-venous anastomoses. On average, 4.3 (2-7 per person) anastomoses were performed per patient. Surgical procedures lasted an average of 6.5 h (5.5-8.5 h) with minimal blood loss (less than 50 ml). Patients resumed normal activity within 6 hours postoperatively and were discharged after an average of 4.1 d (3.5-5.0 d). Postoperative complications included one case each of aspiration pneumonia, lower limb venous thrombosis, and transient delirium, all of whom resolved without long-term effects. Clinical symptoms, including memory decline, mood swings, and anxiety, showed varying degrees of improvement. Patients reported enhanced quality of life, emotional stability, and social engagement, confirming the procedure’s safety and potential cognitive benefits. At one month postoperatively, the MMSE scores of the four patients increased by an average of 0.8 points compared to preoperative levels. Additionally, the two patients who completed the ADAS-Cog assessments showed a decrease in their scores (reduced by 1.0 points and 11.3 points, respectively, compared to preoperative scores), indicating a certain degree of improvement in cognitive function during this period. The ADCS-ADL and NPI scores of four patients varied significantly, without showing any clear pattern.Conclusion:Lymphovenous anastomosis of the deep cervical lymph node-venous anastomosis may provide a new surgical intervention approach for AD, but further large-scale studies and long-term follow-up are needed to validate its safety and effectiveness.
7.Compatibility Study of Biapenem with Parenteral Nutrition Solution Mimicking Y-type Infusion Pathway
Liguo WANG ; Yantao YUE ; Yuanpei LI ; Lijie LOU ; Liang ZHAO
Herald of Medicine 2025;44(9):1490-1495
Objective To study the physical and chemical stability of biapenem and parenteral nutrient solutions when they are used together through a Y-type infusion pathway,and to evaluate the rationality and feasibility of clinical compatibility.Methods Based on the actual clinical infusion rates of biapenem solution and parenteral nutrition solution in ICU,under room temperature and light conditions,simulate a Y-shaped pathway to mix biapenem solution and four types of parenteral nutrition solutions in three volume ratios(1∶1,2∶1,3∶1),and collect the compatible solutions at 0,1,2,4,and 6 hours,observing the appearance,pH value,osmolality,insoluble particles,Zeta potential,particle size and the change of biapenem content of the compatible solution,to investigate the potential interaction between them.Results Within 6 hours,the appearance,pH value,osmotic pressure,insoluble particles,particle size,and Zeta potential did not significantly change.Compared with zero time,the content of the biapenem fluctuated between 93.68%and 100.86%,and there was no impurity peak interference in the chromatogram.Conclusion The physico-chemical properties of biapenem were stable within 6 hours under the condition of room temperature and no light exposure through Y-type infusion pathway and parenteral nutrient solutions.
8.Correlation between type 2 diabetes and hepatitis B virus infection
Wu LIU ; Tianshuo ZHAO ; Liguo YANG ; Genxia GAO ; Wenli MIAO ; Tongxia ZENG ; Yonglan CHEN ; Lina ZHANG ; Xiaoyan CHE ; Lihong WANG ; Junhan ZHANG ; Bingfeng HAN ; Fuqiang CUI
Chinese Journal of Epidemiology 2025;46(5):833-839
Objective:To analyze the latent prevalence of hepatitis B and type 2 diabetes and their correlation through an observational study.Methods:This study used a case-control design. The cases with diabetes were recruited through the diabetes management system and village doctors, while the controls without diabetes were screened from volunteers recruited by village health clinics. Capillary blood samples were collected from the study participants for the measurement of real-time blood glucose level, and venous blood samples were taken from them for the detections of HBV serological markers. Firth logistic regression model was used to fit the relationship between HBsAg positive status and diabetes status.Results:The study included 1 218 diabetes patients, 62 patients with impaired fasting glucose and 491 cases without diabetes. In the cases without diagnosis of diabetes, 11.15% had impaired fasting blood glucose and 4.43% had diabetes. Among those who reported no or unknown diagnosis of hepatitis B, 1.73% were positive for HBsAg, while 18.80% were positive for both HBV core antibody and surface antibody, indicating latent infection of hepatitis B virus. In the non-diabetes group, 0.81% reported hepatitis B history, and in the diabetes group, 2.76% reported hepatitis B history. After adjustment, the HBsAg positive rate was higher in the diabetes group ( OR=2.90, 95% CI: 1.21-6.91). Conclusions:Both diabetes and hepatitis B exhibited a high degree of latent prevalence. The HBsAg positive rate was significantly higher in those with diabetes than in those without diabetes, indicating a potential correlation. These findings highlighted the importance of strengthened screening and management of comorbidities.
9.Study on the immunogenicity and persistence of different types of hepatitis B vaccines in diabetic patients
Wu LIU ; Tianshuo ZHAO ; Liguo YANG ; Genxia GAO ; Wenli MIAO ; Xiaoyan CHE ; Lihong WANG ; Junhan ZHANG ; Fuqiang CUI
Chinese Journal of Epidemiology 2025;46(8):1409-1416
Objective:Exploration of the immunogenicity and persistence of three different immunization regimens of hepatitis B vaccines in diabetic patients.Methods:Participants with diabetes and non-diabetic individuals were recruited from study sites and assigned to different vaccination regimens: the diabetic group (①D60Yeast0-1: received 60 μg Saccharomyces cerevisiae-derived recombinant HBV vaccine on a 0-1-month schedule; ②D20Yeast0-1-6: received 20 μg Saccharomyces cerevisiae-derived recombinant HBV vaccine on a 0-1-6-month schedule; ③D20CHO0-1-6: received 20 μg Chinese hamster ovary (CHO) cell-derived recombinant HBV vaccine on a 0-1-6-month schedule) and the non-diabetic group (ND20Yeast0-1-6: non-diabetic individuals received 20 μg Saccharomyces cerevisiae-derived recombinant HBV vaccine on a 0-1-6-month schedule). Venous blood samples were collected at 1,12, and 48 months post-full vaccination to measure anti-HBs levels. Differences in immunogenicity between diabetic and non-diabetic groups, as well as among diabetic subgroups, were analyzed.Results:This study enrolled a total of 564 subjects. In the D20CHO0-1-6 group, the seroconversion rate decreased from 90.72% (95% CI: 84.84%-96.60%) at 1 month to 74.23% (95% CI: 65.37%-83.08%) at 48 months, and the antibody geometric mean concentration (GMC) decreased from 676.08 (95% CI: 389.05- 1 148.20) mIU/ml at 1 month to 33.11 (95% CI: 23.44-46.77) mIU/ml at 48 months. In the D20Yeast0-1-6 group, the seroconversion rate declined from 93.81% (95% CI: 89.29%-98.32%) at 1 month to 63.72% (95% CI: 54.71%-72.72%) at 48 months, with antibody GMC dropping from 630.96 (95% CI: 407.40-954.99) mIU/ml to 25.70 (95% CI: 17.78-38.02) mIU/ml over the same period. For the D60Yeast0-1 group, seroconversion rate fell from 82.03% (95% CI: 75.29%-88.77%) to 56.25% (95% CI: 47.54%-64.96%), and antibody GMC decreased from 81.28 (95% CI: 51.29-128.82) mIU/ml to 15.49 (95% CI: 11.75-20.89) mIU/ml between 1 and 48 months. The ND20Yeast0-1-6 group (non-diabetic control) exhibited a higher initial seroconversion rate of 97.56% (95% CI: 94.80%- 100.00%) at 1 month, but it still declined to 76.42% (95% CI: 68.82%-84.03%) at 48 months, with antibody GMC decreasing from 1 318.30 (95% CI: 912.01- 1 905.50) mIU/ml to 34.67 (95% CI: 25.12-47.86) mIU/ml. Multivariate analysis on factors influencing the GMC of antibodies revealed statistically significant differences in antibody GMC between the D20Yeast0-1-6 group and ND20Yeast0-1-6 group at 12 months (a OR=0.73, 95% CI: 0.58-0.93) and 48 months (a OR=0.79, 95% CI: 0.63-0.99) post-vaccination (all P<0.05). As for the diabetic population, when compared with the D20Yeast0-1-6 group, the D60Yeast0-1 group also showed statistically significant differences in antibody GMC at 12 months (a OR=0.57, 95% CI: 0.44-0.74) and 48 months (a OR=0.60, 95% CI: 0.47-0.76)(all P<0.05). Conclusions:The seroconversion rate and antibody GMC gradually decreased over time (1, 12, and 48 months) in the four groups. Diabetic patients showed poor immunogenicity and persistence to hepatitis B vaccines. The immunogenicity and persistence of hepatitis B vaccination in diabetic patients were associated with vaccine type, antigen dose, and vaccination regimen. The CHO cell-recombinant hepatitis B vaccine demonstrated better performance in terms of immunogenicity and persistence among the diabetic population.
10.High-throughput single-microbe RNA sequencing reveals adaptive state heterogeneity and host-phage activity associations in human gut microbiome.
Yifei SHEN ; Qinghong QIAN ; Liguo DING ; Wenxin QU ; Tianyu ZHANG ; Mengdi SONG ; Yingjuan HUANG ; Mengting WANG ; Ziye XU ; Jiaye CHEN ; Ling DONG ; Hongyu CHEN ; Enhui SHEN ; Shufa ZHENG ; Yu CHEN ; Jiong LIU ; Longjiang FAN ; Yongcheng WANG
Protein & Cell 2025;16(3):211-226
Microbial communities such as those residing in the human gut are highly diverse and complex, and many with important implications for health and diseases. The effects and functions of these microbial communities are determined not only by their species compositions and diversities but also by the dynamic intra- and inter-cellular states at the transcriptional level. Powerful and scalable technologies capable of acquiring single-microbe-resolution RNA sequencing information in order to achieve a comprehensive understanding of complex microbial communities together with their hosts are therefore utterly needed. Here we report the development and utilization of a droplet-based smRNA-seq (single-microbe RNA sequencing) method capable of identifying large species varieties in human samples, which we name smRandom-seq2. Together with a triple-module computational pipeline designed for the bacteria and bacteriophage sequencing data by smRandom-seq2 in four human gut samples, we established a single-cell level bacterial transcriptional landscape of human gut microbiome, which included 29,742 single microbes and 329 unique species. Distinct adaptive response states among species in Prevotella and Roseburia genera and intrinsic adaptive strategy heterogeneity in Phascolarctobacterium succinatutens were uncovered. Additionally, we identified hundreds of novel host-phage transcriptional activity associations in the human gut microbiome. Our results indicated that smRandom-seq2 is a high-throughput and high-resolution smRNA-seq technique that is highly adaptable to complex microbial communities in real-world situations and promises new perspectives in the understanding of human microbiomes.
Humans
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Gastrointestinal Microbiome/genetics*
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Bacteriophages/physiology*
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High-Throughput Nucleotide Sequencing
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Sequence Analysis, RNA/methods*
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Bacteria/virology*

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